ID GT422598; SV 1; linear; mRNA; EST; INV; 699 BP. XX AC GT422598; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-699 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d1e9e3908a97ba2af4cd52daad5a2bcf. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 12. XX FH Key Location/Qualifiers FH FT source 1..699 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 699 BP; 209 A; 157 C; 155 G; 178 T; 0 other; gccccaacgt tcaagtgtgt gcttgttgga gatggtggca ctggaaaaac cacctttgtg 60 aaacggccca tgactggaga attcgagaaa aagtacgttg ccaccctggg agtcgaagtg 120 caccctttgg tgttccacac caacagaggc cccattcggt tcaatttttg ggacacagct 180 ggccaagaga aattcggagg actgcgagat ggttactcca ttcagggcca atgtgccatt 240 ttcatgtttg atgtcccctc tcgagtaacc tacaaaaatg tccccaactg gcatagggat 300 ttggtcagag tttgtgaaaa cattccaatt gtgctttgtg gcaacaaagt cgccattaag 360 gcccggaaag ttaaggccaa aagcatcgtg tttcccagga aaaagaacct ccagtattac 420 gacatttttg caaagtttaa ttacaatttg gaaaagcctt ttttgtggct ggccagaaaa 480 ttgattggag accccaattt ggaatttgtc gttatgccag ccctcgtacc cccagagatc 540 caaatggatc cccaatggca gcagcaaatc gagaaggccc ttgttgaggc ccaaaaccct 600 cccttccctg acgacgatga agatttttaa attaaccctg ttgtatgtaa ataactttta 660 taaagacttg ttaagtagtc caaaaaaaaa aaaaaaaaa 699 // ID GT422599; SV 1; linear; mRNA; EST; INV; 551 BP. XX AC GT422599; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-551 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 21a697781935bffa6ecbf5181488fdfc. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 13. XX FH Key Location/Qualifiers FH FT source 1..551 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 551 BP; 189 A; 107 C; 132 G; 123 T; 0 other; gggggacatt ccttttctat cccgtcgtca atctgtagct ggaggaattt agtagaatat 60 tttaaatccc gtttcgctta gctaatcggc tgcaaaatgg caatccgacc agtgtacagg 120 ccggagatca taaaaaagag gacaaagaaa ttcattagac atcaatccga ccgatatggc 180 aaacttaaga ggaattggcg taaaccaaag ggtatcgaca acagggtgag gaggcgtttc 240 aagggacagt atttgatgcc caatattggt tatggttctg ccgccaagac ccgtcatatg 300 ctgcccacag ggttcaggaa agtgctggtc cacaatctta aggaacttga agttctgatg 360 atgcaaaacc gtaaatactg cgctgagatc gcccacggtg tctcagctaa gaagaggaaa 420 gacattgttg aaagggcaca acagttaagt atcagggtaa ctaatgggca cgccaggtta 480 cgcagccaag aaaacgagta gatcttgaga ctttggttaa taaaaaaaca acctaaaaaa 540 aaaaaaaaaa a 551 // ID GT422600; SV 1; linear; mRNA; EST; INV; 827 BP. XX AC GT422600; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J15 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-827 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3085a65fbb3b0906f39a680079d8ed03. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 15. XX FH Key Location/Qualifiers FH FT source 1..827 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 827 BP; 278 A; 157 C; 182 G; 210 T; 0 other; ttgaaattaa gaaaatgggc gacgaatact tttgctacgt tactaaatgc aaaaatccca 60 aggcctgcac aatccttctg cgcggcgcca gtaaagatat tctgaacgaa acggaaagaa 120 acttgcaaga cgctctgcaa gtggccagaa atattgtgct gtctcctcgc ctggtgccag 180 gaggaggagc aatcgaaatg gctatcgctc aaagattatt gcagaatgcc actcatggac 240 cttacagagc tttggctcat gctttggaaa ttattccgcg aacattggct cagaactgtg 300 gtgcaaacac aatcaaaacg ttgactgctt tgagggcaaa acacgctaat cataccgatg 360 cggaaactcc ttgcacatgg ggaatagatg gcgaatctgg cgagctggtg gaacagaagg 420 aaaagggact ttgggagccg ttggcagtga aactccagac ctacaagaca gccatcgaga 480 ccgcgatttt gttgttgcgt atcgatgata tcgtttcggg ctccaagaag aaggacaagg 540 agggcgccac tccaagccaa atggccaatc aagaggctgc aggcatgcag gaataaattt 600 aaagtatttt tgcattttgt attttgcaac attttaatta gcttgataat taatatctaa 660 gtaacttagg tagacgcatt ttgtagttca tttacgcgat tacttaagtt cttacatcac 720 tgttaaatat tttattagta agatatttag ttctgatgga aaacgccaat aaaaaaacta 780 aactgttttt ctaactcaaa aaaaaaaaag aaaaaaaaaa aaaaaaa 827 // ID GT422601; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422601; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 154c4dffd1142cfef973a371fba5e0f2. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 16. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 290 A; 139 C; 134 G; 253 T; 0 other; ataacatcca aagcaatcac ttcagaaatt tggaatcgat gagacacgaa gctttcactc 60 aaagcattga tattagttca tgtgtctatt cggcaattca ccaaattcac actcaatctc 120 aatcttccat tgaagaaatt gataataaat tggacagtat caatgaggta tttgacaatt 180 ataccacagt acacttcgtg gaaccttcaa agactagcct ggcaatgacc tacacaattc 240 gagaaagaat ctaccaatgc cctttcaact ttaccgattt ttcaccgttt tatctttgcc 300 ttacaacggc tgccgtactg aacgatattg taatgcttaa cgaatttgca actaactata 360 acgataccac gtctaaagtc aaagcactca tcacggaatt tgaatctgca gtcttgtttt 420 ggcaagcaaa gaacgtggaa gagttcaaga caacagcaca gtccatatat gatgattact 480 ttgtacactg tgctgccgat aaaggatatg ttgtataaaa tattgagtga atttgaacac 540 aagtcttaaa gtggatgttt aagctttaat agcttttagt tcttcgaata tttcgttatg 600 taaatagtat tatgtatgta taacagtcag cctttgttct agaaggtata atttatttaa 660 aaatttgcat tcgcattaac caggtaacat tgtaaaggaa gcacccggtt tgttaatcaa 720 aagtgatcgt gattgtattg ataagtatta ttatgtatat gtattatgag caatatctaa 780 taaaatgtaa aaaaccgaga aaaaaaaaaa aaaaaa 816 // ID GT422602; SV 1; linear; mRNA; EST; INV; 891 BP. XX AC GT422602; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J17 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J17 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-891 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a7903f20e1756edace8da81e9ace462a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 17. XX FH Key Location/Qualifiers FH FT source 1..891 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J17" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 891 BP; 334 A; 134 C; 149 G; 274 T; 0 other; ccaagagcaa aaaattgaag tagccgacat taaatcaaaa ctgacgaact gtggacaacc 60 aggccatcat ggactgggga atgtgaaaaa taccagcaca gttaatagat tgaccgatgc 120 ctcaaagttc acaggaacgc acaaacagag atttgatgaa gccggaaaag gcaaaggcat 180 tgctggacga aaagaccttg ttgattcatc cggttatgtc agtgggtatc aaaacaaaga 240 cagctatgag aaaaaagaat aaaagaaaaa ccatatttat taaaaaatat atgcttaagt 300 ttacaccgtg cactatactc gagaaatgag taacttatat tcaaatattt gtactcgtgt 360 ctgcttccaa tggagcttcc agttttcctt tgcacaagag tttgtttaaa tagaacgttt 420 ttgatatgtc aagtacttat ttagataatt ataagcggtt gtactcgttt agagattttt 480 gcgtagtttt agttttatat tttttcacga tcgtttaaaa atgttaaact cactagacac 540 ttacaattaa ggtacgctgt gctggaatgt gggacattct tttaaataaa actgcacact 600 tcttccagtc tatttcatta aaagtctatg tatcatataa tatgatagta aatgtgatcg 660 tacaaaagtc tagtgcaatg ttgctctaac tgaattggtg taaactaaat caaaagctat 720 atattttatg tataccacgt ctatttcgaa ctccaaactt aattagcata aggtaaagaa 780 taatattttt actttccttc tcgtatgtag aatgtgaaat ttatattaat ttattaaaag 840 acaataaaat aaacaaacaa atgatttgtt caaagaaaaa aaaaaaaaaa a 891 // ID GT422603; SV 1; linear; mRNA; EST; INV; 504 BP. XX AC GT422603; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-504 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0bece336912d0f79383bfe0531076524. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 18. XX FH Key Location/Qualifiers FH FT source 1..504 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 504 BP; 187 A; 76 C; 95 G; 145 T; 1 other; acttgataaa aaggtatttg tcgtcagata tgagcgtttc aaggaaaaat ataatggtga 60 aatggaagtc tacaagagaa cgaagtaatc atcaatccat caaaagtgca ttatgtttga 120 tttaatataa actatgggat atctgttatt tcattttagt gcaacggttg atttataact 180 gcagtcagga gcggatccag tcaagtagta gaaattcaga cttttgttgt gtatgaaatt 240 agaaaaaata acgttagtcg ctgtaaaccc agcataaggt agnttcaagg gggggggggt 300 catgaccccc cctggatcca cccttgattg cagtttaact ccaatatgac tttagattta 360 taataccttg cttctggctt tgccaactta tagctgacta cgatgtttgt gattagatca 420 ataagatacg agttcaccat ctaacaatgt aataaaaaaa aaatgtacaa acaaacaaat 480 ttcaagaaaa aaaaaaaaaa aaaa 504 // ID GT422604; SV 1; linear; mRNA; EST; INV; 841 BP. XX AC GT422604; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-841 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d1225428d2482c5e77621fc4e8ef4d83. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 19. XX FH Key Location/Qualifiers FH FT source 1..841 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 841 BP; 249 A; 180 C; 209 G; 203 T; 0 other; cgaccgactg aacaaggaga tcacattcaa gggacatggg ggttcagtgg atcagttgtg 60 ctggcaccag actaatccag acctgctaag cacagccagt ggcgataaat cagtcagaat 120 ttgggacacc cgtgttcaga aatgtgtggc caccatcaac accaaggggg aaaacattaa 180 tattacctgg tcacctaatg gtaacgcgat tgcagttggc aacaaagaag acttggtcac 240 tttcatcgac gccaggacgc acaagatcga ggcagaggaa cagttccatt ttgaggtgaa 300 cgagattagc tggaataata ccagcgacct attttttctg acgaatggac agggttgcat 360 acacattttg agttatcccg acttgaagag gcagcatatc ctgaaagcgc acccaggcac 420 gtgtatttgc attgaatttg acccaacggg gaagtatttt gccactggaa gcgctgacgc 480 cctcgtttct ctatgggata tcaatgagct ggcttgcaaa agggttttta ccagaatgga 540 ttggccagtg agaaccatct cattcagtta cgacggccag ttgctggcct cagcttcaga 600 ggatttgctg attgatatcg gttttgtaga gacaggggac aaaatcgccg atatcagcgt 660 ggaagcagcc acatttaccg tcgcttggca tccctctcag tatttacttg cgtatgcttg 720 cgacgacaaa gacacttatg ataggaaaag agacgcgggg agtctcaaag tatggggatt 780 tccttcagac taacttgatt tttctacttc aataaacagt taaggaaaaa aaaaaaaaaa 840 a 841 // ID GT422605; SV 1; linear; mRNA; EST; INV; 894 BP. XX AC GT422605; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-894 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9b0208a7b684231ea3015ff3bd52f799. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 20. XX FH Key Location/Qualifiers FH FT source 1..894 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 894 BP; 278 A; 189 C; 193 G; 234 T; 0 other; agtgggccta ctgttgattt caagaggatc cagatgaggc atttcagatt ctggaaaaat 60 ttggactcta cgagtccgcc ccgctttcca ttaaaaccgt gctggaatct cgagaaatgc 120 aatcaaatca tggaggcaac gaaacggcct gcacgccatg ccagaactct aataatagct 180 cgacggaaaa atcgtttcca aatggaaaat tcaaccacgt ttccaatttc ataattggga 240 ctaatcgaat tgccacagat gcagccaaaa gtctcgccat ggcttggcac tggcaaacca 300 ttatcgtttc ccataaggtg gaggaagacg tgaacaggct ggcagaaatt tacgttcaat 360 tggcttactt gatcattgat ggtaagcttc tgaaaatcaa agaactgctt gtaggcctcc 420 cgtttgaatt agaggaaaac gctgttcgag acattttagg gctgcacaga agcaggaaaa 480 tgctactgat ttttgccggc gagccgactg tgcgggtaac gggaactgga aaagggggca 540 gaaaccaaca attagctttg gccttttccc tacaacttaa taaacaggca acgcaagcca 600 caaattctgg aagcattttt tttgctgagc gccggaacag atggaattga tggtccgacc 660 gatgcggccg gcgccatcgt atcctctgga ttatcggcga ccttcaccag tctagatatt 720 aatcccgaag actatttaaa taataacgat tcctattctt tttttaatca gcgccttaac 780 ggagattgtc tggttaaaat aggccattct ggtaccaatg tcatggattt gcactttcta 840 ttaattgaac ctattaatta ataaatgatc aattttaaaa aaaaaaaaaa aaaa 894 // ID GT422606; SV 1; linear; mRNA; EST; INV; 871 BP. XX AC GT422606; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-871 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e2a1cc414aac7b50dd75dd1e1f27aed7. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 21. XX FH Key Location/Qualifiers FH FT source 1..871 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 871 BP; 308 A; 158 C; 171 G; 234 T; 0 other; taacaatacc atgttagaaa atttggaatt atgggataac aaaaaacggc cacttacgat 60 tgctgaatca tttcaagcaa taacattcta caatgttaac ggaaacacaa gtgaggcggc 120 ggatatcgaa gtattgcttt cggggcctcc aagtaatggc cccacccacg gagaaatgta 180 cacaaatgca tcatacacgg ctgtatttga tgttttgaac acatatacgg acattagtct 240 gcctttaaca ttattgcaac cgaaatctgt tggagaaatc actttagcat cagcagatcc 300 cagagatttt ccttcgatta acccgaagta tttttctgat tccgacgact tagaaacgtt 360 gtacaaagca gtggaaattg ttagaaattt agaaaacacg acagcatttc aacgtttggg 420 aatcgaaccc atcattttag atcttccaga ttgtgatgca gattatgaaa agctttcaaa 480 agactggtgg atttgcagca ttcaatatct tacaactgcc ggtcttcatg cggtgggaac 540 caccagaatg ggaaataact cggaaacttc cgtagtgagt tcagaactca aagttcatgg 600 aatcagcaat ttacgagtgg tagacgctgg agtaattcct tatcctattt cgggccacac 660 gagtacaccc gtaatgatgg tcggggaaaa agctgccgag ttaattaaaa atgaatatgc 720 ataaaaatgt taaatatatc ttcggaaagt ccactttctg ggtgacaact ataacacaaa 780 aataatagga aattttgttc gaacaattaa attaaatgta gcgatcttgg gttgaaaata 840 atagaacttc agataaaaaa aaaaaaaaaa a 871 // ID GT422607; SV 1; linear; mRNA; EST; INV; 876 BP. XX AC GT422607; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-876 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0f98ebda7bdc36eb4d2f5cfc8b8ef5c7. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 22. XX FH Key Location/Qualifiers FH FT source 1..876 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 876 BP; 268 A; 112 C; 165 G; 331 T; 0 other; cagaatttag atgtatcttt taataagatt cgtatgggta gtgttgaaga aaaagggggt 60 gatcagtttc tgtcgcagtg tgcgtttttt ttcagttggg ttacaagcag tagttgatta 120 tgttttatat tatatttttg tttttcaagg gaatttgatg tttttgtgaa cacacatttt 180 ggtattcgtt attaagccaa tgttcgataa gattttatta gtggcgttca aaacgtgtgg 240 gaagtgcaaa atattgacgt gatgcaagca attcgattga tatcattttt cgcacttcta 300 aatgttacga gctaattttt ctttaattaa aaaaattgta aagattggtt ggctagatgc 360 tttttcagct tctgggtaat aaggtcattg tggaaggctg ttcccccttt ataatcgggc 420 gtaaaatgag tttctgtttg caaagttatt ctagacgaac cccaaaacgg agtttagttt 480 gtatttcata tgatttggtg aggtgtatgc tacacatcta tgctcagtta atctaaaatg 540 tctcaattgt attttaacat attatttacg caagactgct aattttaaaa acgagcttac 600 taattgaagt aactgaagtg aaaatcacac gactttcagg agccgtcata ttaggcatta 660 catttttagg atagttcaca ttttactaga attaaatagt ttaattttat attgtaactg 720 ttgcgttttt ttttcttgtc acatttaatt agcattagga attgaaatta ccttctcctc 780 gcacacagtg atgtacatgt taagcttaca gttgtgtgca tgtacagata cttaatggta 840 attaataaaa ctatgtatta aaaaaaaaaa aaaaaa 876 // ID GT422608; SV 1; linear; mRNA; EST; INV; 710 BP. XX AC GT422608; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_J24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_J24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-710 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e7dac0995dc2ed707ffc7b1c134d3ea1. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: J column: 24. XX FH Key Location/Qualifiers FH FT source 1..710 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_J24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 710 BP; 229 A; 169 C; 184 G; 128 T; 0 other; tcatagagta agttagttat aatgaatcgc ggtagaggcg gcggccgagg tgggctgagc 60 aggtctttca acaaggaaca gttgagtgcc ttgggggtgg ccaacaacga gatgcccatc 120 ctcgtgacgc aaccgccttc agtttacccc ttaatggagc ggcgccccgt gcctctcacg 180 caatctgaag acacggaata cctattgctg ctacggcaag actttataga tcgcatgcaa 240 ctatcggccg cctatctgaa aatgccctcc aaagaaaaaa cacagcccaa tcaggagatt 300 gatcggctgg tagcgcagct gcccagcgcc aaagacaagt acgattggag cctgctgcct 360 gccgaactgc ggccaaagtt gctcgcccgg aaagtcaaag aagtgccagt cgccaccaaa 420 tccatcaacg tacaggaaaa gctgaagact ctggagaatc tcgaagagac caaggacaaa 480 aaccctgtaa aggggcaggt gaagggtggc gaagaggatc tactggaaga agcctcagag 540 gacgaacaaa tggatgatgg cactgattac gccgacaact acttcgacac tggggaggac 600 tatgaagagg aggacgacaa catggatgaa ggccccattt actaacaccc catgtgatat 660 atctagatat agtaataaaa atcactattt ggaaaaaaaa aaaaaaaaaa 710 // ID GT422609; SV 1; linear; mRNA; EST; INV; 789 BP. XX AC GT422609; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-789 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ede0cc1f9d94442caf5850f513b5392e. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 1. XX FH Key Location/Qualifiers FH FT source 1..789 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 789 BP; 236 A; 161 C; 165 G; 227 T; 0 other; gggggtcatt agcattgcca ttgtttgttg ctgcttcaac acaaccgagt tggttttaat 60 ttaaagtaaa gcagccaaaa aacgtgcaaa aaccatgtcc tatcagttgt accgtaacac 120 aacgctgggg catacgcttc aagaaagtct cgacgaactt ctgcagtatg ggcaaattac 180 gccccaattg gcctgcaaag tcctcctgca gttcgacaag tcgataaacc aaatgctggc 240 cacccgagtc aaatcccgac taactttcaa ggcaggaaag ctgaatacgt acaggttttg 300 tgacaacgtg tggaccttca tgctgaacga tgtggagttc cgcgaagtgc acgaaatcac 360 caagattgat aaagtgaaaa ttgtcgcctg tgatggaaaa agtattgatc cttgataggt 420 ttattggggc cgtgtattac tttctgctta gttgggctac cacttcatgt gcaaatcttc 480 acactagttg ctttgctttt atgtaaatga taaggtgttt ggctccatgt gaatctcacg 540 cttcaaccaa tcgatttatt ttagatgttg ctgaagaggg atgctcgaag aagtagaccg 600 gaatactgag gccccaacca caagttccac catgtacata cttagaataa gcttccttta 660 tcgtcctcat atgttatatt ctgttacaga aatttgtata cctgtagtaa tagtgtgaag 720 ttgcattgtc cttgaaactg tctgcctagt tattaaatga gtacatttac actaaaaaaa 780 aaaaaaaaa 789 // ID GT422610; SV 1; linear; mRNA; EST; INV; 825 BP. XX AC GT422610; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K02 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-825 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c680c77332829eb00fb880cdcb32e18f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 2. XX FH Key Location/Qualifiers FH FT source 1..825 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 825 BP; 269 A; 150 C; 159 G; 247 T; 0 other; gaattggttg caactgatcc agtaatggcc ttccaaaaag gtaaagagtt gtggaccagc 60 ttgagtaaag acgaaccaga aaatcaggct cgtccagaaa acgttgagca actaattgtg 120 ttgctgaccc gtgaatctgc tagacgtatg gttcatttgg ttaattttac cacctcagtt 180 gcaaagaaag ttaatcgaaa tgtaactcgc tctgtagctg tgttggtgca caagtttatt 240 ggagtagctg attcagtagt taagactgct cacttggaac aagtgcaaca agccacaact 300 tctgttatca gaacgcacgc ctattcagtt acattgctgt tgaagcagct aaatactcag 360 ctgacgaata ttttggataa ctgggcaaaa cagttagcta ttcccaccga ggataatacc 420 gaggaaccga agccggaaat tttagaaaaa ctgcatcaaa ctcaaacagt cttaccagtg 480 ccccaaatta aattccaaca gatcaaatcc ttccatgtga acgctataag gaacaaaaat 540 ggtttcgaat ataccatggc agaaaatttg aacaaagatt attaaccctt ttgtctttct 600 gcttgcttcc ttggtatttt tgtaattagt ttagtaatct ttaatatgta acagtgtgaa 660 cgtagccaaa tgataatcgg tttggatggg ttttgcttat tgcttattgc aagttataca 720 ggaccttatt agtagatagt ttccccttct gatttatact actaaatgtg cccttattat 780 cgttgagaaa attatatgat gtttgtcaaa aaaaaaaaaa aaaaa 825 // ID GT422611; SV 1; linear; mRNA; EST; INV; 781 BP. XX AC GT422611; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-781 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 19b97f827f3a9c73d453d828a0c59641. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 3. XX FH Key Location/Qualifiers FH FT source 1..781 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 781 BP; 221 A; 183 C; 180 G; 197 T; 0 other; ttgttactcg aagttcttta gcagccgccc caaagggcaa cttgttattg gtgggacaca 60 ccgccaccct ggacacctgc tcgcgagaac tggtaggcaa gggggcgcgc tcgggcagcg 120 agctagtgaa aatcatccaa aaaatcccgt attgctcgct gatcgaagtg acgggcgtcg 180 gtgagagctg gagcatagtg gaaccgccct gtctacctct cacgcactcc accaatcagc 240 gctttgactg gaaagtcctg ctccaatagg cggtcagcac cagacttttc aattctattt 300 ggttcgcatt cgccactgca tcagggtaaa ttgcgcacat tttttccggt agcagtggag 360 ccgtcgcggt atctaagaac gagggcgcgc ttgtacaata gcgatgggaa caatcgggtt 420 gtttgattaa attaatttag agagaaccaa taggggtatt aagacagatg agggcgatgg 480 aaacctacgc catacaaaaa caaacttttg tacaaaccgc tcttctttta cttgattgaa 540 tatcgctgtt taaagtaggc gattgcaaaa gcagtccgat tattcccgtc gctagtggaa 600 actttcagcc cgatctttat tccagtcgcc agatgacagc gttattggca ctgatacaca 660 ccctccttta aagtaacgat ttttaatcga acgaatgtgt tcataaaagg catcttctat 720 tccaataaaa attcatatga cggcttttcc cgtcctagca gtgaaaaaaa aaaaaaaaaa 780 a 781 // ID GT422612; SV 1; linear; mRNA; EST; INV; 820 BP. XX AC GT422612; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K04 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-820 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; b528431f8ed5fca9e56d6fe0c2a7f7ed. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 4. XX FH Key Location/Qualifiers FH FT source 1..820 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 820 BP; 261 A; 167 C; 201 G; 191 T; 0 other; gccaacaagg acctggaaga aaaggacaag gctctggttg ctgctgaatc tgacgtcgct 60 gcccagaaca ggaagctcca gctgattgag gaagatcttg agcgctctga agagcgtctc 120 gccactgcca ccaccaagct ggctgaagct tcgcaggctg ctgatgaaag ctcacgtatg 180 tgcaaggtat tggagaaccg ctcccaacaa gatgaggagc gtatggacca gctgaccaac 240 caattgaaag aggcccgtct tttggctgag gatgccgaca acaaatctga cgaagtttcc 300 cgtaagctgg ccttcgttga agacgagctt gaagtagctg aagatcgtgt aaaaggtggt 360 gatgccaaga tcatggaatt ggaagaagaa ttgaaggtcg tcggcaattc tctgaaatct 420 ttggaagtgt ccgaagaaaa ggccaatcaa agagtggaag aattcaagaa acaactgaaa 480 tccttgactg tcaagctaaa ggaagccgaa gctcgtgccg agttcgccga aaagactgtg 540 aagaagttgc agaaggaggt tgacaggctt gaagatgaat tgggtatcaa caaggacaga 600 tacaagtccc tcgccgacga gatggactcc actttcgccg aattggccgg ttactagagc 660 acatcactca attttcctat tttaatatcc ttaaagaaaa attgtgtata ttgtattttt 720 gatgtaaact tactattact ttgttctaat gcaccatcac tactatttat acgtttaaga 780 tcctgtgtaa taaaaccgtt tcgaaaaaaa aaaaaaaaaa 820 // ID GT422613; SV 1; linear; mRNA; EST; INV; 859 BP. XX AC GT422613; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-859 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c4ce2e4a39df7327ebbbcac06a8d6c7f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 5. XX FH Key Location/Qualifiers FH FT source 1..859 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 859 BP; 330 A; 124 C; 138 G; 267 T; 0 other; aaaagagcat atatattcag aaagtttcca tgattaatga ataagcataa tgtttgaaat 60 actatgtcag ttagaatata tagattattc tcattcaaaa cagtgtatca gctacataga 120 ctcaacagat aaaatagccg caaataaaac accactaatc aaaaagtatc gcgatatttt 180 aagagagatg caggcttgaa aatgttgtaa taatacactg ttttatagtc gcattagtta 240 gcaacatgta cgtattgatc tatattataa gagcaccaat ccattctgca ccacacaata 300 cagaatgtta tcggaacatt tagaagtcta catctttcaa caataatgtc aagtaccaaa 360 tatatggagt cctctagctg taatgcgaat aagctagaat tgttactaat atatttccaa 420 ggatgacgtg actatgtaaa gctttttgtc ttattgacaa aaattagatt ttttttagta 480 agaactaggt taatctatag aaaatctgaa cgctaaaaat gttagtgtta attaggaatc 540 ggttttccac cctgaaagtt atcaaatcgc ggttttttta gcatagttaa gtcatcagct 600 taagtgttac tacctattga agttacaatt cctttggaaa ctattagtat aaagtttcaa 660 gatcactaac gtaattatga aatgatagga agctttttta tcagtggttg ccgtatagat 720 agataatttg caataaagct gtaacggaat aatcagtcta agaaaaaacg tgttaccagt 780 taaaaggtct actaagagaa aatgttaggt gcacagctgt taaatacacc aagtgcttat 840 caaaaaaaaa aaaaaaaaa 859 // ID GT422614; SV 1; linear; mRNA; EST; INV; 836 BP. XX AC GT422614; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K06 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-836 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 16e5bf34b19acc7a3ab90a5b06352469. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 6. XX FH Key Location/Qualifiers FH FT source 1..836 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 836 BP; 310 A; 138 C; 170 G; 218 T; 0 other; tatagttgca tctgacgaaa aatcttggag caacccagta acgaaggtgc gccagatctt 60 gttggtcaca aaaaacctag cagatttcag cgaaacaaag acaaaaatag cggcgtatca 120 agaggaatag attttcaatg cgtttccaat gtcattaatt tcgattttcc tttggatgtg 180 cagtcttata ttcatagagc tggcagaact gcgagaggaa atcatcaggg aagcgtattg 240 tcttttgtga gcatcaaaga aggccctctg ttagataaag ttgaaaagtg tcttaaacaa 300 ggcgaggaca tttcaatatt taggtcctac caattcaagt tagaagaagt ggaggctttt 360 aaatatcgag ccaaagatgc atggagagca gttaccaaaa tcgccgttcg cgaggcaaga 420 ttaaaagaga tcaagcaaga aatccttaac tgtcaaaaat taaagagcta ctttgaagac 480 aatccaacag atcttcaagt gcttcgtcac gacaaagcgc tacatacagt taaagtacag 540 cagcatttga gcgatgttcc tgattatatt gtgccaccaa ctttgaaaaa tcttgctggc 600 ttgaacaaga aaaccaagag aaaacgcgat cacagcaggt cggcggattt gaaagccaaa 660 ttacaggcaa aaaagaaaaa ccctctagca agtatgcagt ttgaaggttt caaaaagaaa 720 aggaagatgc ttagttatta aaagttttct ttttaagtta atgtaatgtt ttttgaaata 780 cagttaaata agttgtcaat attagttttc ttagacataa aaaaaaaaaa aaaaaa 836 // ID GT422615; SV 1; linear; mRNA; EST; INV; 757 BP. XX AC GT422615; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-757 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3c918a66f0152acd4fda28096b6efa0b. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 7. XX FH Key Location/Qualifiers FH FT source 1..757 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 757 BP; 189 A; 202 C; 195 G; 171 T; 0 other; aggacgcatc gttggaggag acgaggccaa tatcagggac ctgccatacc aagtggccct 60 gctacgaaac aacgcccaaa tctgcggagg atccatcatt gcaccaaccg tcatcctcac 120 cgctgctcac tgtgtggagt tttgggttat tcctgatgtt tccatgctga gcattcgggc 180 ggggtcctcg caatggcgct ctggaggcca gcaggtcgct gtccgggaaa tcgttattca 240 cgaattgtac gatggcgata acagtcttga ctacgacatt gccattctgc acttggccga 300 gaggattaac accatcaatg cctcagaagt gcctttggtc aacaatggcg aaatcttcga 360 tgctggaaca tacggtaccg tttccggatg gggcacaacc ttctcaggat ccagcgtttc 420 tcctgaaacg ttgcgacggg tggacgttcc cattgtaacg caaactcgat gcaggatctc 480 ctacggatca gacatcacgt cccgaatgct ctgcgccgga tacaatgagg gaggaaagga 540 cgcctgccaa ggagactctg gtggacctta cgtgatcaac ggccgtttag ccggagtcgt 600 ttcctgggga ttcggctgcg ccactcctct gttccccgga gtttacgcca gcgttgctgg 660 actcagaacc tggatcaccc aacactctgg agtataagtg agcttgacac cttcaactaa 720 ataaagattc gttgcatcca aaaaaaaaaa aaaaaaa 757 // ID GT422616; SV 1; linear; mRNA; EST; INV; 701 BP. XX AC GT422616; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K08 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-701 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 75a25659ac0dff9c354ce041f2403168. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 8. XX FH Key Location/Qualifiers FH FT source 1..701 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 701 BP; 147 A; 172 C; 234 G; 148 T; 0 other; ggggaggtta gatacgtggc tgttgacaat gaggcaacaa atggggctct gggggctgct 60 gggcgcccta ttggtactgg gggtagggca ggcgcgagcc tactacgtga ccgtggacgc 120 gcacgccgag gagtgttttt tcgagcatgt ggacgctggc accaagctgg ggctgagctt 180 ccagatagcc gagggcggct tcctggacat cgacgtgcgg atcgtggggc ccgacgccaa 240 agtgatttac gaggaggagc gcctgacgag cggcaagtac tcgtttgccg cccatacggt 300 cggcacctac agcttttgct tttcgaacaa gatgtccacg atgaccccca aggtggtgat 360 gtttgatgtg gccgtgggcg agccccccaa gccggcggag ggcgagacgg ccaacaagct 420 ggaggatatg attcgggagc tgacagcgtc gctgacggcc gtcaagcagg agcaggacta 480 catgcagctc aggggccgca tccatcgcgc catcaacgag agcaccaaca gccgcgtcgc 540 catttggtcg ggcttcgagg ctgtggtttt ggtggccatg accatgggcc aggtgtttta 600 cctcaagcgc ttttttgagg tgcgccgcgt cgtttagtgg cagttggctg tacgtttatt 660 gccaattaca gtttttttgt attaaaaaaa aaaaaaaaaa a 701 // ID GT422617; SV 1; linear; mRNA; EST; INV; 566 BP. XX AC GT422617; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K09 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-566 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 06fd00cd8d43cd7ad9a5f63f7056e474. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 9. XX FH Key Location/Qualifiers FH FT source 1..566 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 566 BP; 188 A; 113 C; 134 G; 131 T; 0 other; gggggacatt tctcttttat tctttcgtca gactgtggct ggacgaattt tctataatat 60 tttaaatcct ctttcgttcg actaatcggc ccccaaaatg gcaatccgac cagtgtacag 120 gccggaaatc atcaaaaagc ggacaaagaa attcattagg catcaatccg accgatatgg 180 caaacttaag aggaattggc gtaaacccaa gggtatcgac aacagagtga ggaggcgttt 240 caagggacag tatttgatgc ccaatattgg ttatggttca gcagccaaaa ccaggcacat 300 gctgcccaca gggttcagga aagtgctggt ccacaatctt aaggaacttg aggttctgat 360 gatgcaaaac cgtaaatact gtgcggagat cgcccatggt gtctcagcca agaagaggaa 420 agacatcgtc gaaagggcac aacagttaag tatcagggtt actaatggtc acgccaggtt 480 acgcagccaa gaacacgagt agaactaagt ggttctgatc gggttcttgt ttaatagaaa 540 aacaagttaa aaaaaaaaaa aaaaaa 566 // ID GT422618; SV 1; linear; mRNA; EST; INV; 866 BP. XX AC GT422618; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K10 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-866 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 181dbd92cc8f9ec2e2aacdec3d32257d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 10. XX FH Key Location/Qualifiers FH FT source 1..866 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 866 BP; 259 A; 171 C; 158 G; 278 T; 0 other; atgcccgaaa cactcttgcc atgatccagg aaggtattgc ttgctgcgga gccgatggtc 60 caggcgacta cttaactttg caacagccaa ttcccacgga atgtcgagac actgttaccg 120 gaaatccatt tttccatgga tgcgtggatg aattaacctg gttctttgaa gccaaatgcg 180 cctggattgc cgccttggcc atgcttgtcg cctttattac ggttatgaat gtggtcgttt 240 ctattgtcct catccaagca ctgaggaaag aagaagaaca agctgacacc taccggaagt 300 agaatcccaa aatcagccct attgtacaat tttatttaca tacataccgt tctcatgctt 360 tgcaataagt ctcacatcag tgcaacaagg ctgatgcccg aattgctgtg ctagaaagag 420 agcaaacata aacaaaccaa cgttaggggc ggggtttcag taaacctgga ctccgccctc 480 aacagcaatt gcctctcttt ctaacaaatt gctttattgc actactttgt tccgctactt 540 ttcgcgattt gtttgatagg acatatacaa attttcgact tttttagtac aatattcgtt 600 tttaagtagt ttatcttagt gtagttttaa cccgccttca attttgatat gttatatagt 660 attttattgg tagtttgtag attatttttt aaatatttaa taagatcaaa cagggtttgc 720 ttgctatgtc aaaaataaca ttttctttta aatattcatg taccgttttc cacaaagttt 780 gactttaata ttcaatggca aagttgtaag cgctgaatgg taacgaaatt atacataatt 840 attacaggaa aaaaaaaaaa aaaaaa 866 // ID GT422619; SV 1; linear; mRNA; EST; INV; 860 BP. XX AC GT422619; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-860 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2f8f959b217349a999e99b961454d825. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 11. XX FH Key Location/Qualifiers FH FT source 1..860 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 860 BP; 263 A; 155 C; 188 G; 254 T; 0 other; caggtacatt gaagagtctt tagggcaagc cgaagaagaa aatatccaag tgaacgagaa 60 agtgccttgg agggccatat taacttctct accatattgg gcggttgtat ttggagctat 120 tggcgaatct tggggaagca cgtttttgat tacagaaatc cccacgtatt tgagcaaaac 180 cactgatatt aacattgaaa aaaatggact ttactcgtca gctccttacg tggtagctgc 240 ggtgcttaca gtaatctacg ggccattagc agattatttg attcacagca atgtgacttc 300 tagaaaaacc tccagacgca tatttcatgg cttgggagcc tttctgcccg cattagccct 360 agtatggctg gcatatgaga aaaaccgatg gggaatagca gctttactta tagtggcaat 420 cagcttgaat ggggccatgt tttgtggata caacgtgaac catattgata tatcgcctcg 480 cttttctgga gtgctttttg gtatttcgaa cggaattgga caaacgctgg cgattttagc 540 acctctactc gtacagttta ttgtgtatga cgagaccgat aaagatttgt ggaggacaat 600 gttcgttata gctgctatta tttatgcagc aacagctgtg gttttcataa cttatttgtc 660 cgctgaacga cagtggtggg atgccatttc caggccaagc aagggagatt tccagtcgga 720 attacaatct catcgtgatc aggaattaat taatcagtaa ttgtatttta tgtacctaac 780 attagcttac aaaatggact gtaattcatg tttatcaata aaataaaata attaacttga 840 aaacaaaaaa aaaaaaaaaa 860 // ID GT422620; SV 1; linear; mRNA; EST; INV; 709 BP. XX AC GT422620; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-709 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f92d846db0c61c4a7a0d97918df0b24e. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 12. XX FH Key Location/Qualifiers FH FT source 1..709 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 709 BP; 257 A; 122 C; 160 G; 170 T; 0 other; gggggacaac aaaaagcagt tgaggttata aaatagatag gcagaattaa aaatgtttgg 60 cgaaaaagcc ggtaacttaa taaaagaact atccagatgt gaaaacgtat tgccgccata 120 caacaccgac atggtgaagg aagtttgtgc cgaaataaag cagcttgatg acctgaacag 180 gagcaatgga gcgattgttg ccaatgaaag cggcttggca gtcaacagca accccttata 240 tcccactctc agaattgggg taacagccct gaaaagaaac gtccgctgtc tgatcgctta 300 ccaccataac agactcagga tactcagaac catgagatgg gagtttggta gtgttttacc 360 ggcagatatt aaggtcaaca tgagctcaga agaagtcgag tggttttgcc aatattccag 420 aaatttggcc aaatacatga gatcaattgg cgaagatggt ctaaatttgg ggctggatct 480 tcgacctcct aaatcactct acattgaagt gagatgttta gtggattacg ggaagtttga 540 gttatccgat ggaacggtgc tattgcttaa aaaagacagc agacactatt tgccaagaac 600 tgagtgcgag gagttgatta caaaaggtat ttttcaacat atcttataat tatcagtgta 660 aatacaactc tgtaaaagtt aaaaaaaaaa caaaaaaaaa aaaaaaaaa 709 // ID GT422621; SV 1; linear; mRNA; EST; INV; 832 BP. XX AC GT422621; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-832 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a670e8106f12e902eac1112721387a66. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 13. XX FH Key Location/Qualifiers FH FT source 1..832 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 832 BP; 226 A; 194 C; 195 G; 217 T; 0 other; tttggggtac gacagtccgg cgaaacatcc tcgaatcggc gtagccgaaa agcactacat 60 tatggaggct ttggaacatt gcgaagagga aatgagcgac tccaacttgc cgataaggca 120 aattttggct tcgcttccgt tttgggcgct cgccatttgc aacatcggca acacctgggg 180 aatcactctg ttcgacttgg aggtccccac ctatctccag aaggtcctgg gcttcgacat 240 taagacgaac ggcattattt catccttgcc ccaaatcacc cagctgggac tttcgctggt 300 gtttgcccca gtaaccgact acattgtgga gaaaaaggtc atctctctga tgaactgccg 360 cagactattc caagttttag gttccttggt gccggcagcc accctagtgt ggctggcatt 420 catcagcaaa tcccaggcaa cactgataat catcctactg aacgttataa tcggtttaac 480 agtcttcttg tacaatggct cctatataaa ccatgtggat atttccccaa aatacgccgg 540 actgatgctg gggatggaaa atagtatatc gcaaatggtg ggattaactg ggcccatttt 600 cgtgcaatac atggtgccag atttgagcga cattctcctt tggaggaacg tgtttcttct 660 agtgtctgtg ataaccgcta ctactgcgac attctttatg attttcggat cggccaagat 720 ccagtggtgg aactcatgga gtgcttcgaa tgcagaggcc gagcaatgca ctttgagcgc 780 accgattata gaaaataaag tatgatatat ggacaaaaaa aaaaaaaaaa aa 832 // ID GT422622; SV 1; linear; mRNA; EST; INV; 826 BP. XX AC GT422622; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-826 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 008f41f1bbe4b20b29db5e8566853c5d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 14. XX FH Key Location/Qualifiers FH FT source 1..826 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 826 BP; 246 A; 164 C; 182 G; 234 T; 0 other; aaaccccaga ggtaaaacac gaaaggttgt tcatagtatc cgtcctgggg ctccttgtaa 60 acttagtggg tgtttacgcc ttccaacatg gacattctca tggaggaggt tcatcgcatg 120 ggcacagtca tggaggtgcc tcacatggtc attcccacaa taatcattct catgaccact 180 ctattgagat ggacaatgag ggtcctaatt cccaaatcat gaaaggtgta tttctacata 240 tattggctga taccctggga agtgtgggcg taatcatatc agcagtgcta atgcagatgt 300 ttggttggat gatagccgat ccaatctgtt caatgtttat agcggttctg atcgctctca 360 gtgtcttggc tctaatcaaa gactctgtgg ccgtcctcat gcaaagacag ccgtattcct 420 tggacagtgt actgccacag tgctaccaaa aagtgatcag cttagctgga gtgtattccg 480 tgcaagaacc acacttttgg acgctatgca gcaacgtata cgtgggtgct atcaagctgg 540 aagtttcgaa aaatgtggac ccgaaatacg tggttcaaca cacgcaaatg atttttgctt 600 ctgtcggtgt taaacagttg tatgtgcaat tagactacac acccatgtga ctgtagtctt 660 tgggtcgcaa gtgatttttg caaattatat aaagcaaatc gtgatcgttt tgattaaaat 720 gtgtttacat tgtttgccga aagtaaattg gaaaataatg cgaacaaatc aaatttctac 780 gtaattacca ataaaactgc aagtcacgga aaaaaaaaaa aaaaaa 826 // ID GT422623; SV 1; linear; mRNA; EST; INV; 760 BP. XX AC GT422623; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K15 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-760 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 42c2b0235c39c452b0c23e5ed2077cbd. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 15. XX FH Key Location/Qualifiers FH FT source 1..760 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 760 BP; 265 A; 127 C; 125 G; 243 T; 0 other; aaaaggggcg gtgctaagca agtttagcct gtattttcac gatatcttgg ctaatcaaac 60 cactcccact gatatgcggc atatttgctc caaactacat catgattact atcaaaagat 120 ggtaatattc cagagaaagc acgaagctgc ttgtgtgata ttactttctg ataatcaagt 180 taattgtgaa actgctgatt acgatagttt tcctataata gtttcctatc ctccaagaag 240 tcctccccaa ctagatataa tgttaaaaat gatatcagaa actaatgagt ttttcagtct 300 caatccggta actaagtttc gctatcagga tcaatgtacg tactgtttaa ccatggtgga 360 accgaacata tactttgtga ttttgttcga atcgaagaaa actgaaaaag acgcctggat 420 tggcaatttc gttaacgact tttgcaataa ccttcgatgt acaaaaatat tcgtcagttt 480 gaaaaatcct gtaaagtaac tggccgtaaa tattatattt atgtaaacca aacgtgaaag 540 cttttttcca acccataaag aagaaaaaat ggttaaaagg ggatttttag tgaccccttt 600 ttagtatcaa accaattttc agctgtttcg tttcagagag aaatccttta agtgcgcttt 660 agaaaaataa tttattcctg gaattcatag ccagaaattc tttaatgtgt ttttgggatt 720 taaatataat ttttttgaaa acaaaaaaaa aaaaaaaaaa 760 // ID GT422624; SV 1; linear; mRNA; EST; INV; 764 BP. XX AC GT422624; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-764 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 16f001ce054c7340887051c75298f24f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 16. XX FH Key Location/Qualifiers FH FT source 1..764 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 764 BP; 245 A; 146 C; 157 G; 216 T; 0 other; gggggttttg tttgtagtta tgaggaacct gcaggtgcct ttcaacaatg gcgtttgaac 60 tggtactttt aggggcgctg atcttcattt tcatcgtctc attctgtgga ttagtgcaaa 120 aaaatcaaac agacttacac cagagagcgt cttttggcag aacgtctggc aagacgggca 180 gaacgacaag cccgacgaag atccgtggat tttacggata tttatgtgaa tccctcattc 240 aattccccgg caaatggagg gatttttcct ggctcgcctc ctccttactc gcaacattat 300 actgaaccac cgccaaaata cgaggaagcc atcaaagtgt gtgcggcaaa tgtccaaaca 360 agtcctcaac atgtggaaac ccgttcgagt gaaactcggc caggttccga accgcctcca 420 tatacgatca ctttaacaac gccaattgcg gccacatcaa tacgaaatta gtagaattta 480 actttataac tggtgctcaa tgaaagcttc acagattctt aagatcttag tggcaagagt 540 agaattagga aagagtaaat taagggctta attgaggaaa ctagtaatta aattaatttg 600 ttagttaaat aattcgtatt taaatacata tactcattaa ggtatttctt gagggaaaag 660 agagaccggg aactttcggc tggttcgaat ttttctggat tatgttcaat tgcattttat 720 ttaaaaaaat tatattaacg tctgcacaaa aaaaaaaaaa aaaa 764 // ID GT422625; SV 1; linear; mRNA; EST; INV; 826 BP. XX AC GT422625; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-826 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ee8fb32894a37afde2e29003fd8fcd1d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 18. XX FH Key Location/Qualifiers FH FT source 1..826 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 826 BP; 261 A; 136 C; 193 G; 236 T; 0 other; ataaaataca aataccatta caagcgtttt ctaaataaaa tattgcttgc attttctgac 60 cattttattc taagttccgg agagttcaaa aacagcaatg tctttgcatg gatctaatga 120 ttggatgaat gctgctctta caactgggac atctattatg gcagcggagt ttgaaggggg 180 cgtagttatt ggggccgatt ccagaacaac tactggggca tatattgcta atcgggtaac 240 tgataagtta accaaaataa gtgattatat ttattgctgc cgatctggat cagctgctga 300 tacacaggcg attgctgaca tcgtatctta ccaattatgt ttccatggaa tggaacttgg 360 tgaagaaccg caagtagaag ttggggcgaa tatcttcaag gaattgtgct acaattaccg 420 agattcgctc atggctggca tcctagtagc tggttgggat aagcaaaaag ggggacaagt 480 gtactcaatt ccaataggag gtatgtgcgt aaggcagcct gtttccattg gtggttcagg 540 atccagctac gtttatggat acgtggatgc aaactacaaa agaaatatga ccaaggaaga 600 atgcacccag tttgtactga aaactttggc tctggcaatg gctcgcgatg gatcatcagg 660 aggtgtggtg cgtctaggaa tcattactga gaaaggagtt gagcgaaagg ttatactggg 720 gaatgaattg cccaaattct ttcaagaata attggagaat ctttgtataa taaagattgc 780 ttcatttttt taatacacta tgtcatcatc aaaaaaaaaa aaaaaa 826 // ID GT422626; SV 1; linear; mRNA; EST; INV; 676 BP. XX AC GT422626; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-676 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a14f123d1a0f99a2902b3484340e8b43. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 19. XX FH Key Location/Qualifiers FH FT source 1..676 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 676 BP; 207 A; 149 C; 175 G; 145 T; 0 other; ggggaggaga attcagcgta agggacctct ggtcgtttat catcaagatc aaggtctacg 60 cagagctttc cgcaatatcc cgggtattga tttaatcagt atcgagaagt tgaatctact 120 gaaattggcg cctggcggtc atgttgggcg tttcgtcatc tggacggaat ccgctttcca 180 aaagctggac aaagtgttcg gaagttggaa agtggcttcc agccacaaaa agggctacaa 240 tctgcccgaa actaaaatgt ccaacaccga tctgtctcgc ctgttgaagg ctgatgaaat 300 caaggcggtc ctgcgtcgtc cacagaagaa gattgtgcgt cgtgtgcgac gcctcaatcc 360 attgacgaac gccaaggcta tgttgcggct caacccgtac tctgcggttt tgaagcgcga 420 agcaatccta tcaggccaga aaagaagctt ggctagagag gaacttcttg ccaagaagcg 480 cgggatcact ttaccggaaa caagtccggt aatcaggtcg gccaagctgc aggctaaaag 540 gagggtccaa atcttgaagg cccagaaggc aaaggctgcc aagccgaaaa aagttgccgc 600 gaaggcacca gctaagaaat aaatatgatg tttcttagtt acgaaataat tatcaagtaa 660 aaaaaaaaaa aaaaaa 676 // ID GT422627; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422627; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 36b514f3d2a4dc34bea50e19f6e00d76. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 20. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 264 A; 120 C; 154 G; 278 T; 0 other; ctgacaatta tttgttgtta catgctcagc tctttatgtt ttatgatttt tcctgtttat 60 aacaataact taacagaaaa ttaacaaaat ggcatgagtt aaaccacacc tggctgctca 120 agacataatc aaagtagtaa aacacttgtg ttgctggttt agtgacgtgg atgtgatcat 180 ttaggttata tttactccac tttcccaaag aaaaactgga tctttaatag aaaatcaaaa 240 tgttcgaaat aaccgacaca caaaaaatag gtgtaggcct agctggattt ggggttttct 300 tcctattttt gggaatgtta ctactatttg ataaaagttt actggcatta ggcaatatcc 360 tttttattgc tggattggca gcagttattg gtagagagag aacattcaga ttcttcttcc 420 agaggcataa aataagagga agccttgcat tttttggagg tattatcata gtgttatttg 480 gttggcccat ggtgggaatg atttttgaag cttatggatt ctttatgtta ttcagtggtt 540 ttttcccagt tgccataaac ttcttgagga gggttccaat cttaggcaca tttttgaatt 600 tgcctggtat tagtaaaata gttgataaac tggctggaga ctccaaccga acaacggtct 660 aactaactaa tatgtatgca tgtgaaatac aatgattctc gtctgtaatc taaacactaa 720 cttaataagt atatggtgta tgtacagaaa caactttttc acttactgtg ttggaaataa 780 aacatatgtt taagtaggaa aaaaaaaaaa aaaaaa 816 // ID GT422628; SV 1; linear; mRNA; EST; INV; 825 BP. XX AC GT422628; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-825 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; de7e9d0434e3885cb1906503e37d5421. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 21. XX FH Key Location/Qualifiers FH FT source 1..825 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 825 BP; 259 A; 166 C; 206 G; 194 T; 0 other; ggctcactga actgccagag gatttcggaa gtctgtgcaa gctccgatac ttagacctgt 60 acaaaaacaa attggaaaga ctgcctttga gcttcagcca actggttggc ttgaagttcc 120 tcgacttaaa ggacaaccct ctcatcccaa ctattgccaa agtagccggt ccttgcgtcg 180 acaataaagg gtgccaacaa tgcgccaagg atgtggtaca tttcttcaaa cagctggagc 240 aagaggtgga gtcgcgtaag caggtcagac agaaaatgct agaaatcaac cagcaggaga 300 aacaggcaga gaagaaaagg ctgaagaaag agcgacaaag caagacagca attaaggagg 360 ttcaagtggt gaaaaacagc acagcagctg aagtgcctca gttggttcca gctaatagca 420 aaaagccttt gaagcctctc gcaggttcag caaaaagccc gttgttttcc tggtttgtct 480 attcgatttt tctgctgctg gtgctgctgt ggttcttgtt ctcttggagg gtgtcgttgg 540 cgcaaagttt agcacctaaa atcggcgagg tttatgaaaa cctcctggac cagttgccta 600 gtaattatag ggcgttagcc cagtcttttg gggattcaat catagcccta caggtgggca 660 cacggaacat atcaaggcag tgtgcagatg ttgtgtacaa cagtgaaata gtgcagactg 720 tgttgattaa agtgtacaat ttaattggga agaacactag ttagtcccct agtgcaaggg 780 cgaataaatg ctttaactac taaaaaaaga aaaaaaaaaa aaaaa 825 // ID GT422629; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422629; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 20d15475de25f81accf6b5c022214a8c. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 22. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 309 A; 116 C; 113 G; 278 T; 0 other; attagatgga tgatctccat gaaggtacgt gcaaagttta ccaaaatact aaaatgatca 60 actcaaatgg actaacgcta gtgtaattgg caacaagccc tttttaaggt agtttttcca 120 ttctgtgtaa tcgaacttat taatttcaag taataagttg ctatagccct tcctgctagc 180 gaaacgtcag gaagtaattg ttttctctga cagcctaaaa gcctgaaaaa ctcatcatta 240 cccaattaac tgtttattct tcataaaaaa aaatctacta tggatataag tctaaattgt 300 agaaagttat cttttattta catctagaca acagtttagt tagcaacatc cattgaatta 360 ttttaattta gtgttgcaaa agtttcacaa catactgttt tcaacatgca caagcgctgt 420 aattcaaact gcaaagcaaa ctaataatgt atacataaaa taattgattt tggcctcgaa 480 agctttaaat atttaagtag accagaagtt gaccattgca cattcagcca gagcatagcg 540 taaattttca aatagatatt taggatttgg ttttaggaac attttatgtg ttaccacaat 600 taaattaatg agtataatat ttaagccaaa tgtatttgaa ataacagact ggtcattgta 660 aatagatttt agcattaatt tctagatttt gtaatagcaa ttagtttaat atattgccca 720 aactagtaaa tacattaaat gatttaataa atattcttgt gttagaattc ataatataaa 780 tatataatta catgatttgt aaaaaaaaaa aaaaaa 816 // ID GT422630; SV 1; linear; mRNA; EST; INV; 809 BP. XX AC GT422630; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-809 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 69421b8c43401763c525f068af18489a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 23. XX FH Key Location/Qualifiers FH FT source 1..809 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 809 BP; 191 A; 225 C; 231 G; 162 T; 0 other; tgcaggatgt tccgcaggct gcccaggagc tggcggcgct gttttggccc caacaggacc 60 tgcaggggct gcagcccctg ctggaggcgt ttgtgcaccg tgaagtatac gattgggtgt 120 tcggcctgat caagccgacg ttggctgcag aggactggcg gctgtatggg ctgtccaggg 180 agttttgcca tcgcaatggc agtccgaagc aactgggggc cacgttctat aactttgtgc 240 ccaatgctgc tgtggtggag ctgagcacgc tgcccactaa acggacgcca ctggaaaaac 300 tccactgcct ccagagctcg tacgactaca ttttcgccga agtgaaggga gccctcattt 360 ccgtcattgc caaatactca gacaaagaac tcgagctgcc gctgatcagc aacgaggagg 420 ccgcgcccat cctgatggcg gtgatcatga agtcgaaact tttctacgcc cacagcgagt 480 tggcctacat ccgcaccctt gggcggggcc tgctggaaag ccacgcccac ctcaggcaaa 540 tcttcgcgaa gttcgaagag gcgctggagc gcatccagcg ccagccaatc agtgacgcga 600 tgggctacca gctggccaat cagatggacg tatgcgagac catcgagttt gtgcagcgat 660 cagcggccaa gcagccgaaa aagacgattt ttgctgaaga aacggggcgc gtcgcccgct 720 tgatcacggc cgccaccact gaaaccctca atttttgatg tgttttttct ggacaataaa 780 accggtttta tcgaaaaaaa aaaaaaaaa 809 // ID GT422631; SV 1; linear; mRNA; EST; INV; 783 BP. XX AC GT422631; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_K24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_K24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-783 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f7b17e83b3f7eb71d403dd875e66e7e3. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: K column: 24. XX FH Key Location/Qualifiers FH FT source 1..783 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_K24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 783 BP; 276 A; 129 C; 124 G; 254 T; 0 other; caaaaaaagt ttgcagttac ttaagaaaaa atccaagccc aataagacca acacagaata 60 gggatacgag tgtcttcact ctgaaaaata cgcctcgtca agtgaaggca ctgtacctct 120 aggaactgga aaaatatcaa cggtagacga aggtagtgag cacatatcta gatcacagat 180 agctttacca gaacaccaat tagaaaaagc tcacacttca aaaactgttg ataaacctat 240 ttccagttca caaaatcgga taattgaggc tgaaactacg gtagaaaaaa acaaacctga 300 aacagaaagg gactcaaatt caatgctgaa caagttccga aatattttta agttttaaat 360 tcagattgca agtttgttca aagtgaagac caggtctgaa atgtgtgtat tgcgtttact 420 atcttaactt acatattttt cttgtagcgt ttcatattac gtttctaagt aagagtgcgt 480 ctcattttct tttctttggt ttattttgtt agcttatttg aagtaagtta tgtctgatca 540 aacgggtgca gtttttaccg aaacaaatcg ttcattttct ttttaatatt cttttgtcac 600 tcatttctga agacatgaat atcacgtcta tgtaagaaaa ataccgctac gatttctgtt 660 ttttttttag ttaacttaag tatcgagtat attaataata gtagaaatga atttgttact 720 actccgactg ttattatatt tttgttttaa tagaatcccc tacccaaaaa aaaaaaaaaa 780 aaa 783 // ID GT422632; SV 1; linear; mRNA; EST; INV; 837 BP. XX AC GT422632; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-837 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 7da668c79ff205766e9302abaf94ca92. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 1. XX FH Key Location/Qualifiers FH FT source 1..837 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 837 BP; 277 A; 151 C; 201 G; 208 T; 0 other; atcagagctc gtcaacagga ggcagagtcg gagatacgaa atctcttcga caccctgcac 60 ctattagtag aagctgccaa gggaagcttg aattcaacca tcgaagaaac ccataagttc 120 tatcaggaag agttggaaga aatcaaggct gaagctctag caaatggcaa aaatatttcg 180 cgctgcattg aactggctga tgcagttctg ctagacctgg atgaacaagt gaactccaca 240 gcaagtgaaa cctatgaaaa tcttgagaaa actgctgaag aagcagtgaa tcgggcgctc 300 caaatggcgg tcaccgcttt gcaagagctg gaaatgttaa acggaaaagt ggaaacgtgc 360 aaagatgatg aatgtgccac tgagctggat gtggaaatag tggaattcta cgaggaaatc 420 gttgcagatc tggacaacgc catcacgttg gctgaagatg cagttttcat aaaactcact 480 gccgagctgc agaatagcac tgagacaatc gactactaca acgaacaagt tcaggagctg 540 attgaagatc tgaaaagttg cgctgaatga tttgcgtttg attgttcaaa ttgcggcttc 600 tagatgtttc cagctcagtt ttaccatcag tttaataatt ttttgggacc caatggagta 660 gtggcaatgt tcaaattggc attgaatcta caatgagttt gcttctcgaa tgagacaatc 720 gattgttttt tatatgaagt gcttggaacg atcgcaggca gcaattaaga taaccaaatg 780 tagatcgatg tatcggattg ttgattacaa tgagttgtcg aaaaaaaaaa aaaaaaa 837 // ID GT422633; SV 1; linear; mRNA; EST; INV; 813 BP. XX AC GT422633; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L02 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-813 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d1ca4e3a2d4205fe053b6f77ec82ea67. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 2. XX FH Key Location/Qualifiers FH FT source 1..813 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 813 BP; 251 A; 171 C; 202 G; 189 T; 0 other; ctataccagc gaggactttt ccaacctgcc cgatcaatgg gactggaggc tatatggggc 60 tgttaccccc gtgaaagacc aatcggtatg tggttcctgc tggtcatttg gcaccatcgg 120 caccgtagaa ggtgcattct ttttgaaaaa cggaggccat ctagtgcgac tatcgcagca 180 ggctttaatc gactgctctt ggggctttgg aaataatgga tgcgatggag gagaagactt 240 cagggcctac caatggatta aaaagcatgg tggcattcca actgaagagg attatgggcc 300 gtatttagga caggatggat attgccatgc cgataaactg cccaaagtag ccaagctgtc 360 cagttggacg aatgtgacca ctaacgatga gaacgcgctc cgactggcca tcttcaaaca 420 cgggcctatt agtgtggcaa ttgatgccag ccagagaacg ttcagcttct attcgaacgg 480 cgtctacttc gaggagaaat gtcacaacaa agtggatgag ctggatcatg cagtgctggc 540 cgtgggctat ggaagcatta atggaaacca ctactggctg gtgaagaaca gctggtccaa 600 ctactggggt aacgatggat acgttctaat gtcctccaag aacaataact gcggagttat 660 gagcgccccg acttacgtta cattagaata agggactttt aactgatata actgataaag 720 cactgcgtaa actgtagcaa tgtagctccc caaaaatcgt acaaatttaa ggatcctaat 780 aaaaatataa aaattaaaaa aaaaaaaaaa aaa 813 // ID GT422634; SV 1; linear; mRNA; EST; INV; 844 BP. XX AC GT422634; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-844 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; fe4c106f8bb17a533372d29a941bd244. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 3. XX FH Key Location/Qualifiers FH FT source 1..844 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 844 BP; 278 A; 181 C; 171 G; 214 T; 0 other; ccagcagcga ttctcaatat gacattctgg aaggaagaag acgaaaatcc catccacaac 60 gcatcgctga gtggagataa actgatcgcg tggtctgcgg agcaaagtat ccaacgagtc 120 aacactgtga aaaatattaa aaatcgaatc ccaaacactc gattcagcga agtggttctg 180 agctgtgtat cagcgagctt ttataactat tttcaactga aaaaatggaa agttcccccg 240 caaatgtccg tcctaactcc cctgttgttt ctctatcctg aatcaaatag cagcaagcta 300 cccgagctga ggaacgactt tactttcgcc gacttaagcc tgcccttagt agacccagac 360 cactttataa atggactgga ggatgtgaaa gtcagcgtgg agaaaatgag gaagagtcct 420 accactgctg tcatgtactt tttggtgaac tacatatgtg cagtgattcc ttcacagctg 480 ttactgaaga taacaactac caaggcgttt actgccacta tctcgatagt gcctggagcg 540 ggacgaatca aagtgtttgg cgattgcgat cttgaaacta ttatctcaac tgcgccacac 600 tcaaatgaaa caggaatttg ttttaccatc ttgacttatg atgacaaatt ccaaatttgt 660 gcgttcgtgg acaaggcgat aatgtctgaa caagaagacg ttcagtacct tgtcgacgaa 720 gtgtttcgaa acatggacct cctaaacaaa cagagtctgc agacgttcgc atagatttac 780 tgtaaaaata ctctaggaag tcaatttaat tacaaaataa actaagtaaa aaaaaaaaaa 840 aaaa 844 // ID GT422635; SV 1; linear; mRNA; EST; INV; 836 BP. XX AC GT422635; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L04 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-836 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4209d544fa35de07737b6c5b83d3a602. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 4. XX FH Key Location/Qualifiers FH FT source 1..836 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 836 BP; 237 A; 185 C; 216 G; 198 T; 0 other; cgaaacgatt gacgccgcct tggagccctg gaacgaaatt gttgaagctg agggatcggc 60 tgtgatcggg tcctctttgg tgaaactgtc caaaacgggc tgctattaca gcgagtgcaa 120 cttgggcaac tttgtaactg atgcctatgt cttcgcgtac tctcaaaaca ttgtcgatgg 180 ggcctggacc agggcgccca tagccattat caatgctgga gggttgagaa ccgacgtgga 240 aattggcaat attacctacg gcgacatgct aacagcgcaa ccgttctcca acaccgtgga 300 ctttgggcga atagaaggca gatatttgaa ggaggcgctg gaacaggcgg ctggatccta 360 cagcaatgcc cgaattacct ccgacctgaa actccttcaa gtgtctggaa ttcacgtggt 420 ttacaacgtg tccagacctg aaggagaccg agtggttgat gtgaaaatcc gctgccagca 480 gtgcgatgta ccagactatg atgagctgga cttggaggca acctacagta ttgcagtgcc 540 ttctttcctg gttggaggcg gtgacgcttt cacagccata tcggagaaca ttgaaaatgt 600 cgtaactgga caaacggaca tggacatttt caggctctac ttgcagtccc gatcgccagc 660 ttttgctgaa attgagggcc gcattacggt gcttggggac gaagaaatcg aaaccaaggg 720 ctttgatact tgaacaacta gttaatattc ccattcagtt atagcctagg caaacaagcg 780 gtgtggaaaa taaacaatca tttattcaat ttttaaagaa aaaaaaaaaa aaaaaa 836 // ID GT422636; SV 1; linear; mRNA; EST; INV; 824 BP. XX AC GT422636; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-824 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a2d4450b42a4d87ebf36f28512f30803. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 5. XX FH Key Location/Qualifiers FH FT source 1..824 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 824 BP; 268 A; 127 C; 201 G; 228 T; 0 other; tggagaaaca aaaatgtagt gtccaaagtg aaaaatcaaa aaaattgtgg agcttgttgg 60 gcatttgcgg tttctgaaac tattgaatcc atgcaggcga ttaagacgca gcaattaacg 120 gatctgagca ttcaacaatt gattgattgt tcatcgtaca ataatgggtg taaaggaggc 180 gatacttgcg ccttgttgcg ctggattaaa gtcaataata ttgcaatcat gaatgagact 240 gattatccgc tcgtacttga ggaccaaaaa tgccagaaga ctgatatgag tgaaggagtc 300 aaggttggga cctatcaatg taatagtttc gtcggtagag aagacattat cctgaaacta 360 ctggcgatta atggtccagt cgcggtagcc attagtggcg aaacttggca aaactacgtc 420 ggaggtgtga tccagttcca ttgcgaaggc gatttgagcc atgccgtcca aattgttggg 480 tataatttaa cagccaaagt gcccttctat attgtaagaa attcttgggg ggaagatttc 540 ggcgataatg gttatttgta cgtggccatt ggaggtaatg tatgcggatt agcctacgaa 600 gtagcagctg tgactgtgct ttagagtccg gcagtaaagg taaacatgaa ttttgcttta 660 tgtggggttt aagactgggc caaggaacca tttgggaatc gtattttcac attaaacgtg 720 aaaaattact taaactaagt gatgatctcg actatggata agagcctaga tatgtaaata 780 ttaacgaatt attacaagct taaacagtaa aaaaaaaaaa aaaa 824 // ID GT422637; SV 1; linear; mRNA; EST; INV; 833 BP. XX AC GT422637; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L06 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-833 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e9803b7ba82959a5c11902bef4908415. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 6. XX FH Key Location/Qualifiers FH FT source 1..833 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 833 BP; 264 A; 188 C; 180 G; 201 T; 0 other; acccgcagcg gatcgcgcca ggatgtgatc gctatcatta acaccttcca gagaatgggc 60 ttcaacatca gggcgaatga tgttctttca aacggcactg ttgaagacgt cctcgccaaa 120 ttagatgaag ttctccgaga caaagagtcc ctggcagaaa ccaacagctt gttcattttc 180 ttcctcaccc atgggttggc cggggacgaa ttgtacgcta gggatggtat cttgcagtgc 240 aggaaaatct ggacgaaatt catcgactgc aacgagttaa agggcaaacc gaaaatgttc 300 attttccagg cctgcaaagg agactattac gccaaaattg acgatcatcc ctctttcaac 360 tcggaccctt ccactttggt gcacttgaac acttttaagc ccaaacacgt cggaatggat 420 atgctgatct tcttcgcaac catcgaaggc aatgaatcgt accggaatcc cttagaagga 480 acctggctaa ttcaggaact gtgcagaaac ttgtcatcgt atggccgacg ggatgacatg 540 atctccatct gcatgagaac gacaaaatgt gttacgggaa attactacca tcaagaggaa 600 ggtgaaattt tgaagcagat gcccgtcttg ataacaactt tgtcgaaaaa gttctacgtg 660 aatagaaaca aagagaggca cctgctgctt gaagtcctca aagagcaaaa agaaacgaag 720 gatctactag tgcaattggt cgctcttttg caaccgcacg tctaaaatgt tcaacgctat 780 ttcacgtaca tttaaatata tattgtacac gttctaaaaa aaaaaaaaaa aaa 833 // ID GT422638; SV 1; linear; mRNA; EST; INV; 590 BP. XX AC GT422638; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-590 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f627f67b5074a905c92a25053096e9ee. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 7. XX FH Key Location/Qualifiers FH FT source 1..590 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 590 BP; 215 A; 77 C; 97 G; 201 T; 0 other; ggggctgcca acccttggaa ttgttacaga aagtaactgt cgaaactatc ctaaaaattg 60 ttacgattag ttcgataatg ttccccacag atgtgaacta aaagtcggaa agtaattcca 120 atggagcccg gcttataagc cctaaaacct tttcaaccga tattttggcc ggccgtgcaa 180 accgtcatag tttgattata tgctaatagt cagcaaacaa gttttaattt tctccttatc 240 ccttgagagt ttttagattt tgtatatttt ggtgtgttag gtaatagcta gaatcttgga 300 gagttccaaa caaaaatgtg atagtagaaa aaactttgac aagtttttta gtttattaat 360 catataaagt atattatata tattttatga cgtgtttatt aaagttttaa gccttgtatt 420 tttatattta aatattttaa gacagttgta actatatgcc ttatggtaat cctatgaaaa 480 gagatgcgta aatatattaa tgagaaaata acttaaaaat tgatgtgtat gattacataa 540 aaaatatacg tcaacttgtt agtagcaaaa tgaaaaaaaa aaaaaaaaaa 590 // ID GT422639; SV 1; linear; mRNA; EST; INV; 829 BP. XX AC GT422639; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L08 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-829 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ed228d457d7b4b1dc5e45cee2ad32f49. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 8. XX FH Key Location/Qualifiers FH FT source 1..829 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 829 BP; 264 A; 202 C; 206 G; 157 T; 0 other; gcgctggcca acgcgtacga gattctcaaa gacgacgaga gtcgcataga ttacgactac 60 atgctggata atccggatga gtactacgcg cactattacc gctactaccg gcgccgagtg 120 acccccaaag tggacgtgag aatcgtgatc gtagcagcca tatccatcat atcgatcgtc 180 cagtactact cggcctggca gcgctacgat agtgccatca cctactttag tacagtgcca 240 aagtaccgca acagggcaca agacattgcg aagcaggaag gcctgctgcc cgacaactcg 300 aaaaggggcc ccaagcgaca gaaaacttcc aaagaccaga tggaggccat catcaagaag 360 gtcatcgagg acaagatgga tatcaaaggg gcctacgcga agccgaaact atccgacgtt 420 gtctgggtac agctgatcat cttcccctac acggctgcta tctatctcta ctggcacgct 480 tcttggtttt ggcgacacac cgtcctcaag cagccctaca gcgacgagga gaagctctac 540 ataatcagaa agcactttaa gatggggcaa catcaattcg actcacagga ggaggagcgc 600 aagcaggaac tgctcgatag gaagctgtgg gtgaaggaga acttcaatga gtggtttgag 660 gagcaggaag aaatcaccaa gaagcagcta gcggagaata accggcataa acagtacagg 720 cgctacatga aaactaaggg agtcggccgg atgacctttg atgattcata atttattata 780 tttaggaaaa tacagttttt acataaaccg ccaaaaaaaa aaaaaaaaa 829 // ID GT422640; SV 1; linear; mRNA; EST; INV; 850 BP. XX AC GT422640; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L09 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-850 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f97e782ff5042afcbadb47f2bcffb67c. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 9. XX FH Key Location/Qualifiers FH FT source 1..850 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 850 BP; 279 A; 161 C; 180 G; 230 T; 0 other; catacaagca ttcaattcgg ttttccatga cattgtggtt taaatagccc aaaagacgtg 60 gtacaagtca cgaactcaat caacgttgaa ttatttaact ggttgataga cgatgccgca 120 ggagataaaa acgtcgctcc ggccgggcag gaaggacacc atactgatgg attcgatgtg 180 tggaattcca ccaatatcat tatccgagat tcggtcgttt ttaatcaaga cgactgttta 240 gcaattagat gcggggagaa tataactgca tatagtttgc gatgccacgg tagccacgga 300 cttagcattt cagtgggatt cagcgccgac gacattacag tcaatactct gagaaatatc 360 acaattcgag attcgtattt gaactccggc gccaatggaa tacacattaa aacgcacaat 420 gatggtggtc ctggtttgat ctccggagta acttacagca atataacctt taacgacatt 480 gaaaaatttg gtgtacaaat tcagcaaaat tatccgcgag gagatgcagt tggaaatgtt 540 cctattaatg atctaacgtt catcgacgtt ggaggaaatg tggcgaatac cggggtacct 600 gttttcattt tatgtgcaga aggcgcctgc aataactgga tatgggaaaa tgttcaaata 660 gaaggacaaa atgaaaacag ttgcaattac cagccaactg gctttcaatg ctgaccaagc 720 cccctatgtc caaatgcata ttagatattg atgtttattg cgtgttaaac tgataatgtt 780 tccactccag tacagttaca gtttgtacat gtataataaa taatgctaca gctaaaaaaa 840 aaaaaaaaaa 850 // ID GT422641; SV 1; linear; mRNA; EST; INV; 841 BP. XX AC GT422641; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L10 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-841 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 24ab2126712215be77ae8326d2e0043f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 10. XX FH Key Location/Qualifiers FH FT source 1..841 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 841 BP; 252 A; 171 C; 188 G; 230 T; 0 other; cgctgatgcc tgcacaaatc cagcttttta cggatgcagt cgcactggaa ccagtgacaa 60 tgctttgaat ccaatcaaaa ctgccaggat cagaaccgtc gaatcattct catttaagta 120 cggtactgtt gtagtaagag caaaaatgcc agctggcgat tggttgtggc ctgccatatg 180 gctcttacct cgttacaatg catatggaac ttggccttct tctggtgaaa ttgatctttt 240 ggaaagccgc ggaaacaagg attatgtcaa tccaaatgga atgaacgtag gtactcaaca 300 agtcggccat actctccatt gggctccttc tgttggtgca aatcaatggg ccaaaactca 360 tttcgataaa aatgatcctg ctggctacga cactgatttc catatctaca aactagtctg 420 gaactcgaat ggatttatat tttacatcga caatgaggag gtgggcgtca tcaatccccc 480 agagggcgga ttttgggaaa tgggcgaatt cgcaagcacc ggagtggaaa atccctgggt 540 agctggaaca aaaatggccc catttgacca acagttttat ctgattataa acttggctat 600 tggtggtgtc aactattttt cggatgaaaa tacgaacgct ggaggtaaac cttggtctaa 660 cgaatcaccc acagcttcaa gcgacttttg gggtgcaaga agccaatggg aaccctcatg 720 gaatagaaat accgacagtt cacatttgat tgtcgactat gttcaagttt acgctgcgta 780 ggtttactgg ttttttattc attgtaatat aatatttaaa ggtgaaaaaa aaaaaaaaaa 840 a 841 // ID GT422642; SV 1; linear; mRNA; EST; INV; 852 BP. XX AC GT422642; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-852 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9b183b2a30d587a56e1c48af8291852f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 11. XX FH Key Location/Qualifiers FH FT source 1..852 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 852 BP; 255 A; 126 C; 125 G; 346 T; 0 other; atagcgatca tccgttctac ttttttgtgc acaattcagg aattgttctt tttagtggaa 60 gagtagtcgg ctaatttact tattttattt tcattaatta tattatttta cctatattag 120 cgcaactgaa agaaggaaga cattttgggg actaattatt actacaaata aagcattgtt 180 tataaatttt gattattttt taacactcta aaaaggacag gagacacact gagcttcatt 240 ttctggcagt ttttaaaatg ctgtttgctc taagtgaata gaaacatgtc aacttaatca 300 agtgttctat tatgtaaaat ccatgtagtt ttcggatgtg attttcattc agcaaatgcg 360 aaaattattc gcgggtattt atatatgtag atgctccata caaaaagcgc agttagttat 420 tcattataac gtttttatga tttatttctt taagtttact tttcttctta ctttgtctta 480 tcagctatta ttcattactt ctcttttatg gtgtggactt ccttgcaatg catattgcgg 540 tagactcttt gccacttcct gcagaatatt tctttatagc ggaccatcct ttcttctatt 600 atattagaac cggcgatgtt gttcttgtga ctggaagagt ttcctcaaac tagacatgtt 660 attacttatt ttactcttgc aaaccgttct ttcatttaat atatgaaaat tattttttac 720 ttgatttctt tatgctaata gcttgcggat ttttataata ttttgctata ttgtgaaggc 780 ctacatgcat tgggaatata tattattata ctttcacaaa tataatttta aaaagcaaaa 840 aaaaaaaaaa aa 852 // ID GT422643; SV 1; linear; mRNA; EST; INV; 800 BP. XX AC GT422643; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-800 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ed4a46a1dd3c07d7a305307fc2ee2a3c. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 12. XX FH Key Location/Qualifiers FH FT source 1..800 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 800 BP; 253 A; 140 C; 180 G; 227 T; 0 other; ggggattcac aatgaacctc aaggtgtgtg ttgccattct gggaacattt ttagctgtcc 60 aggctagtaa ccaatctcca gccttaacag gtaccactaa ggatgtaatc gttcaagtac 120 tggatgaggt tctagcagta gtgagtgaac gtctggccaa tgcgagcgct gtggttgaca 180 atcttcaaag taatcctgat caaatccaaa ccaaaattgt tgacgaagta caggcagtgg 240 taaaaacgct caatagcgaa gtcgtgaatg ttttaaacaa agtgagaaat aacgcctcag 300 atggaggtaa agcgatcatc gattgcgtat tagctgaaga cgatccagct gaagcaatta 360 ttgaagatgc aacagccagc tttggagcat gcatttctaa tgacaccagt gtagttctgg 420 ctgctattag tgatggtctg gatcaattaa cgcagctggt tgctgatatt caggctcctg 480 tggatacttt aagtagctgc acgagttcag acacgatttg tcttttgagt tttgccacct 540 ctgaagttgc agtactaaaa actgttcctg acttggtcca agagtatatt gctcagtgga 600 aaaaaatata tagtacaatt tctgccgatg caacagaatg tgctgcaagt gcgcgcagtg 660 agtacaagtc gcaaattatg gcagttttcg aagacagtgt tcaatgtatt ttgaatatta 720 aaagcaacta gctcaaagaa tgtttgtgat atttaagtta tttaatacat ttgttttatt 780 acaaaaaaaa aaaaaaaaaa 800 // ID GT422644; SV 1; linear; mRNA; EST; INV; 852 BP. XX AC GT422644; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-852 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5345dfb8ff233606f68a4116efd9c77e. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 13. XX FH Key Location/Qualifiers FH FT source 1..852 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 852 BP; 219 A; 194 C; 216 G; 223 T; 0 other; gactgcaaaa gtgcgctcga gacgcagccc gctgtcaatg tgttcgatgc tcagacggtt 60 ctgagccaga aaaagtcgca ggcagcgatc agtcccagac agccgctggg gctcagacag 120 agcccgaata cgattctgag cccgtgcgaa gcaccgatcg tccaaagcat aacgacgaca 180 gacgcgctga ccccctcaga cagcctccaa tcctaggggc aaaagcggcg cgaaacgaag 240 aaattgagcc tcagctcgaa gaaagtcttc tgaggtgcca gtccgggttt gttttgtttg 300 gtcgacatcg ccccctttga gggcggaact gacgtctagg ggcggcgacc cctcctcaat 360 gtctcactaa tcggaacatc tccaaggctg gcaggaccat tggacctcat aggaggactc 420 ggggttcgcg cccctccctt tttggtacct gttatatgtt gcaatttgtt actctttaag 480 gtgtacatac acgcaaacct tgttgttaag aggcgggtga aggaaacagc gtgtggttcg 540 atgtacggca gtgggccctc tccaacagaa aacgcccaat ttttctcaat ttttgttcaa 600 tttttgtgag ttgatttctg tgatttagtc gttagtgaga attattgctg aatatgtcta 660 gaggccgcag aatggcgcag atggccaggc tggacacatt ttatgcggct gcattttgtt 720 ttctgtctca aaggtttttg taaataggac gcgcgtagct gttaaggatt tttgttaatt 780 agattttata tatgacagat tataattttg agcaaataaa cacgactttt ttcctgaaaa 840 aaaaaaaaaa aa 852 // ID GT422645; SV 1; linear; mRNA; EST; INV; 803 BP. XX AC GT422645; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-803 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 92798bdcb7daad6473164309199cbd21. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 14. XX FH Key Location/Qualifiers FH FT source 1..803 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 803 BP; 244 A; 181 C; 205 G; 173 T; 0 other; cagggcattt gctgaagagg acctggagga ctccagacgt cgcgcgagta gcaggatcaa 60 agagaaccat ctcttcgact ccagaggggc ccgttacttt gacgacgcga tcaatgatga 120 gactgcgtcc actctgaagc agatcagggc ggccaagaaa gtctcctatg tggaagacgc 180 cgatctggaa tcggcctcaa acaacctcag aacgcaacgg atgctggatc ggtcggagaa 240 aatcctggat agcattggca tcaatgagag ctcagcgagc aagagggctt tggcgaacga 300 gttttcctat cagaaaagat cgctcaagat gtcgtatgac gcggatgcgg ataatctgac 360 taaatggaag gcagttcccc cggcggaaaa tggagactcc accaccagtg ccgcttctat 420 cagagctcgc cgctctcggg ccagaatcga cgatattgat gatgaaatgg ccgccatgca 480 ggagaaacaa gctgttagag agcgccgcgt tgcccgccta aagcagctgg ttgcagaatg 540 cgactcagaa accgttgacg acaatggcgt agccaccaca gaaaagaaag tccacttcta 600 aataatgtag ttgtttttaa gctgttgtga caatgtgtat cttaagccca ctagcgaatg 660 aggcagggca ttaatgaatg cattttgggc ataaaacatg ttgcatttta ctctctagtt 720 gtattacagt gtacatacgt ctccaattgt atgatatttt cacgcaaaat aaacgttaaa 780 cggctaaaaa aaaaaaaaaa aaa 803 // ID GT422646; SV 1; linear; mRNA; EST; INV; 795 BP. XX AC GT422646; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-795 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; da2d59e1da0054ad3cd33a57cf43c4fa. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 16. XX FH Key Location/Qualifiers FH FT source 1..795 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 795 BP; 240 A; 132 C; 118 G; 305 T; 0 other; ttttcctaag tccctttcct agttgcaagc cggctactac ctccaaatcc catttggccg 60 cgcagaggga gcaatatatt gctcctgtat tgcttagttc cagtagcatt gaaaattgtg 120 ttatagacag atcttctcct gccctctacg gctcagctag tcaagggatt gtaattcctg 180 aaacaggctc tagtaatgct tgagtattac attattattt cgttggctgt gacgttataa 240 aatttaatta atttcacccg gattgttttc ccaatgcttt aaaaatgatc tgtttgtatt 300 gtaaatattc ctgtaaatta catgaattat ttatagtact ttgtaaccgt actttgcaac 360 taactgtact gtgaatctgt ttttatgcca tgtactactg actatattat tcttttgaaa 420 gtattacctg acttgccatc cagttctata tttaattcta attttatgaa gctgaaaaaa 480 tgtatgctac acgtattttt agtttagttg tttgaacgtt tgttgttaat attttgttta 540 aaactctact gttacgccct gattaatgta tgtttgaatt ttcaaaccca agtgcctata 600 gaaactcatt cttaccaatt tgcaactatt tattctaatt atttaaccct tgtcaatgag 660 agacaatgtt ataaggcata ggaatggtgt cggtattatc aactgtgtaa ataaaaaact 720 tatcatactt aatgtgtatt tatttgtaca tttgttttat aatataaact aatttacaaa 780 aaaaaaaaaa aaaaa 795 // ID GT422647; SV 1; linear; mRNA; EST; INV; 806 BP. XX AC GT422647; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-806 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 409618e68c9df6e8ee3830a21d3d3e76. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 18. XX FH Key Location/Qualifiers FH FT source 1..806 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 806 BP; 249 A; 158 C; 156 G; 243 T; 0 other; ccgcgatgat gttacactag atgaaattag gactatcgag agaaaccgac agccgcccga 60 tgagggcccc atgtgtgaat tgttatggtc cgatccacag ccccaagatg ggcgcgcgcc 120 cagcaaacgg ggcgtaggat gccagttcgg acccgatgtt actcacgcgt ttatcaaact 180 gaataagctg gattacatta ttcgaagcca tgaagtgaag gactcggggt acgaggttgc 240 tcatgagggg cggtgcatca cagtgttttc ggctccaaac tattgtgata ccatgggaaa 300 caagggcgct tttataaatc taactggtga aagtatggtt cctaaataca ttgtctacga 360 cgcggtgcca caccctgacg taaggcccat gatgtactcc accaattcat tgatgtatcg 420 actctgctag tacctgctag tacctgctat ttaagcgtag tctaatttat agtcggatta 480 tcaatcaatc aattttctac acacacatat tagggtaatt gaataattag tgaatattaa 540 aatttcagaa cttaaatttc caatttgttg gtgatgtcaa taagtttgtg ttatacttgc 600 catcgcaagt agctctttat ttacacaaac tgaaaattgc gttttctccc atcattaatt 660 attattatta ttattattat catatagaac ttgcattgaa caattacacg atatatatcc 720 gtttaatttt gatatatcgt cccatttttg atagtgtata ttttaataaa acctagattt 780 aaacccgcaa aaaaaaaaaa aaaaaa 806 // ID GT422648; SV 1; linear; mRNA; EST; INV; 842 BP. XX AC GT422648; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-842 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e5af755183ce11f403abb664c0d7e439. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 19. XX FH Key Location/Qualifiers FH FT source 1..842 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 842 BP; 316 A; 115 C; 150 G; 261 T; 0 other; aaagaaggaa atttgattag aaatgagtct gcgaataata gtatcagaac tgattctcga 60 gctcctagtg ttagaagttt agagatctac agagaacagt actgttgctg cgcaagaaga 120 tcgaaatgcg aaaaatctct actagtcact gtaaccgtat taataattat aataatagtt 180 cttgtcatag ttatagcagt gctagcttca aacaagcagt tgcatcaact cttcagcgta 240 gcaaatagat gaagccgagc aagaattgac gaataattgt tgataaaatc ttgtaaaggg 300 cttcaactaa ttgatacctt tatagtttct tatgccactc gaagaactct ccaaaaaagg 360 gaaatgtgtc tttgcatcaa tattggaatt aatacatata taaatcaatt gaggggcatg 420 ttggcagagt ccattggaag caaacaatat ttcaataagc tactagcaag caaatttata 480 aaatttgatg aaagtcgatt gactcattta tatttctatg ctttggacag cagcatttgc 540 taaacaaagc atttgacctt cgtgatattt acttcaattt ttcccactta aaaaaattga 600 actataaaaa atgtattagt atcattacta aaagcaacgc ataaaattac aaaactttaa 660 ataatttggg gtgtgggatg ggatttggac tggcttggat ttgttttgta ctgtgtacaa 720 atatgtataa gtataagtga caaaacagtt ttgtatgtat tttatagaat gttaaacaat 780 gtgtaggtat ggtttattaa aggtaaccta caactttaac aaagaaaaaa aaaaaaaaaa 840 aa 842 // ID GT422649; SV 1; linear; mRNA; EST; INV; 622 BP. XX AC GT422649; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-622 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5b8889a782ec006eb4d9ca9f059dd2bf. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 20. XX FH Key Location/Qualifiers FH FT source 1..622 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 622 BP; 179 A; 138 C; 157 G; 148 T; 0 other; gggggccatt ttcaaaatgg aagtctttct ttaaggttta gtctgctgtt ggaagttggt 60 gcgctacagt tcttagttaa tgtgagtgat cagaatgttg atgcccaaag agaatcgtgt 120 cgctatcttc gagcacctct tcagagaggg tgtaatggtg gccaaaaagg actatcacgc 180 ccccaagcac atggagtttc aaatcatccc aaactcgcag gtgatcaagg cccttcagtc 240 cttgaaatcc aagggatacg ttaaggaaca gtttgcgtgg agacatttct actggtacct 300 aaccaacaca ggcattgagt acctcagaac tttcttgcac ctgccttctg agatcgttcc 360 ttccacgttg aagaggcaag ccaggacgga aagtgcacgt cccaggcttg ctggagcccc 420 acgaagtgac atgcttccca agggcactga agaccgcact ggatacagga ggcacactgg 480 aggtccagga tcagataaaa aagcggacgt gggcgctgga attggtgagg tcgaatttcg 540 gcccggtttt ggcagaggtc gccctcaata aactttttaa atgcaaataa acacatttat 600 ttggcaaaaa aaaaaaaaaa aa 622 // ID GT422650; SV 1; linear; mRNA; EST; INV; 823 BP. XX AC GT422650; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-823 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0149fe05d9625d7f0a0365721a6eb988. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 22. XX FH Key Location/Qualifiers FH FT source 1..823 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 823 BP; 223 A; 220 C; 166 G; 214 T; 0 other; agttcgaccc cctgcaggag aagcaccaaa agaatggtac tgaggaccaa aagccgcctt 60 caaagccctc tacacctcag aatgagggtt cgaattcgag gccccagtca gcatccatga 120 tttctcagca gtctgcatcc cagcagcagc ctgtgcccac gtcagctgca gcagtgaatt 180 cacacatgaa cgactacttt gccaggagtt ccttgggaat gcagtatcca ggtctgtctg 240 ggtatccaca gcagtacaat tttccgtcta ctggccgata cgctccccaa gtaatggaca 300 gttcccaagt tccaagttca aacgcgtttg caaacccacc ccaaggtagc ataccttatc 360 ccactcagca acaatacggg cacccgccgg gcggcgtagt ctatcccccc ggacaggcca 420 acccttattc taaggccttc acgcagccca acccccagca gcatggatac atgccccctc 480 gtcaataagt ctgttttaat ttcacgaaac aaaataagca cgttctacta ctaggatacg 540 ttatgttgtt aagctgtcgt tttaggggat tgtcttctcg tcatgtttag cactcatttt 600 agaatttgat tttaaacata ccattctgca aatgaaagcc acgttttcgc cgagtgcgcc 660 ttcagcccaa gtggtttcgt tgcaaattcc actaggcaaa tgtggcattc atcgttgaaa 720 tggttctgtt gttgtttttt taatctatgc ctttcacttt gattatcagt acattttgta 780 gcctataagc aataaagtgt gggcagcaaa aaaaaaaaaa aaa 823 // ID GT422651; SV 1; linear; mRNA; EST; INV; 802 BP. XX AC GT422651; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-802 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ff94bed7d2cb8298d62760b245a2e911. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 23. XX FH Key Location/Qualifiers FH FT source 1..802 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 802 BP; 247 A; 167 C; 168 G; 220 T; 0 other; gacaactttg acatcgttga gggtctcatg acgactgtcc acgccacaac tgctacccaa 60 aagactgtag atggtccttc aggaaaattg tggagggatg gtcgtggagc ccaacaaaac 120 atcattcccg catccactgg agctgccaaa gccgtaggta aagtcatccc tgccctcaac 180 ggcaaattga ctggcatggc cttcagagtg cctgttgcca acgtttcggt tgttgatttg 240 actgtcagaa tagccaaagg cgcctcttac gatgacatca aggctaagat caaggaagca 300 gcagaaggac ctcttaaggg aatcttggga tacactgatg aacaggtagt atcgtctgac 360 ttcgtctccg acacgcacag ctctgttttt gatgccgccg ccggaatttc tctgaacaac 420 aacttcgtaa aattgatctc gtggtacgac aacgagtatg gttactcgaa tcgcgtcgtc 480 gacttgatca aatacatcca aactaaagat gcttaaaaag tattaatgcg acactagatt 540 aagtacgtac aaaagttgat aaaaacttta atctcagatg ttcgtagagt gtacagtatg 600 tcgcgctctg tacgttagga tcccagtgta gtcagaagtc agtctaaaaa tacctttatt 660 aactttttga cacgctgcaa aaacttaata agtatatgta aatgtggtga attttgtgtt 720 acttgcaccg tactgtttgt ttaatttctt atttatgtat tcaaaaatat aatgtgtatg 780 ctccaaaaaa aaaaaaaaaa aa 802 // ID GT422652; SV 1; linear; mRNA; EST; INV; 760 BP. XX AC GT422652; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_L24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_L24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-760 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 89073aa98a87205d11249826f60410cd. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: L column: 24. XX FH Key Location/Qualifiers FH FT source 1..760 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_L24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 760 BP; 251 A; 96 C; 132 G; 281 T; 0 other; gacataaaac accctcggta tttgtaacct ttaatgttta aaaattcatt tactacagaa 60 agtattgtta ttgttggtgt tcttttattt ggtttttgaa catccctact aagtaatgta 120 ggacgaacta aatcatctca ttttctaatt agacggagga aggctgtaac atcatcctcc 180 tgattatgag attccaattt ataaagagca ctagcaagtt ggaggaatac acaaaatata 240 ctttttgttg tacttttatg aagaatttga ttcaacattt taaagttgtc acctggtttt 300 acttttttgt tttcatttgc tgttgacgtc gagatagagg gttagttata gaaatcaatt 360 tgacgcttta aagatttgaa ggtgctctac tataatttaa attgttgaat taaatcataa 420 tttatttgag aaataaaggt ttcgataacg agtttgaaga tcatattagt aaggctgcaa 480 agcaatgcac gtttttaaag tattttcttt tggaaatacg gctgttattt tttggttttc 540 tggatcttat tctcatttaa gatcgctgaa ttgtaagaat tgcgtttcct gtgggaaagt 600 cgttagaatt gccaaatatt cagagaaggt atatttaagc gctgcattgt tagtaatttc 660 tgctaaaaac tagttgttaa aaattatttt ctacatttta acatgtgtat gaagaaagac 720 aaaccatgct ttggtcaaac aaaaaaaaga aaaaaaaaaa 760 // ID GT422653; SV 1; linear; mRNA; EST; INV; 801 BP. XX AC GT422653; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-801 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ac0f3803c84db196849c0b7a41940d42. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 1. XX FH Key Location/Qualifiers FH FT source 1..801 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 801 BP; 245 A; 197 C; 161 G; 198 T; 0 other; gcaaacgcta ccgaatgtta taaacaatct tctatataca gagattcgca ctagcgccat 60 gacaaatctg ggaaacattc tgatgggata tttggatgac gaaggttcca cccaagatga 120 ctccaaagtg ggacgtcttt ctctagccaa cgaaaaactt gcatacatgc ccaagttagt 180 gtgcgaagcc tttgggccca tcattaaaat actggacatt tcggacaaca acatcggaaa 240 cttggatttt ttggactatt tcaccgaatt gacctcactg atagccgaca agaatccaat 300 cagctccgct gacaccaaca tcccttgcat gcccaagtta gagctgctct atttgaacaa 360 gtgcaaaatc agcaaccttt gctgggtgga agcgctgcgc tacaactgcc cgaaattgag 420 atttctgtcc ttaatgggca acccggtagt tccagccttt atgactacgg gaaacgtcta 480 ccagtacctg cagtacaggc tctatgtgat atcagcgatc cccaacttgg tgcatctgga 540 cgacaagaga atcactgccg atgaacgggc gcaggccaag aaaatgttcc caactccatt 600 tgtgcaccat ctgctcaagt ccactcaagt gcgggttcct cacctcttaa gaaagctcac 660 gggtagaacc agcaattatt ttcccaaatc ccctgtgagg agccatttac acccatccaa 720 accaatgggg aataactatt tggtctaggg agaatttaat taataaattt gttgttttca 780 ttcgaaaaaa aaaaaaaaaa a 801 // ID GT422654; SV 1; linear; mRNA; EST; INV; 831 BP. XX AC GT422654; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M02 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-831 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e8dfdef5449d1d589202408e9083598f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 2. XX FH Key Location/Qualifiers FH FT source 1..831 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 831 BP; 227 A; 175 C; 166 G; 263 T; 0 other; tccagcaagc ttccggaatt aatgctgtgc tgttttttgc gcaacaaatt ttccaagacg 60 ccggaaccac gctggctcct gcatattgtt ccatgataat tggtggagtt caatttggga 120 ccagttttgt cactcctcta gtctcgaaca tgttcggccg caaagtttta cttatcggat 180 ctgccatcgg catggctctt agcgaaagca ttctcggcat ctacgacatc atcagggcag 240 cagacgaaga caaagtcagt tcactcagct tcctaccaat cgtgtctctg gttctatata 300 ttattacgta taacgttgga tttggtcctt tgccatgggc agttattggc gaaatatttc 360 ccaacagcat caaatcttca gccagcgctt tggcgacttc agtttgctgg ctcaccagct 420 tcataatcac caaatggttt tcacaagtgg cagaggctat tggacaaggg caatgtttcc 480 tcggatttgc cggattttcc ctactagctg cagtgtttgt tttcttcgtg gtgttggaaa 540 caaaagacaa aaccttagca gaaattcagg ttgatttgaa taagtaactg cgatttttta 600 tatagttaat atgatgtgag gtttttgatt cattaaagaa cactcaacag cattctccta 660 ttcaacggaa atttgcgctt gcttcagttt cactgcaagc cactgctcgt ttttacgcgc 720 cgattattca tattaattat gcattgttaa caggattaag ggattcgctg cacaaacttg 780 ttaaaattgt acatattaga ctttttgagt aacaaaaaaa aaaaaaaaaa a 831 // ID GT422655; SV 1; linear; mRNA; EST; INV; 854 BP. XX AC GT422655; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-854 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2bb1d01b4ee0564814c785d2af522e65. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 3. XX FH Key Location/Qualifiers FH FT source 1..854 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 854 BP; 282 A; 162 C; 191 G; 219 T; 0 other; agaaacgcag gaaggagatg aaaagcgaaa agaacaacgt ggaaagtgtg aacgaggatg 60 gaactaagag gaaaatggct tttcttgact tgctgctgaa aagtcgcgac gaacatggac 120 aaccactttc acaggacttt attcgacgtg aagtggacac ttttatgttt gccggccacg 180 atacaacagc ttcagccatt agcttcgtct tgttctgttt agctaatcat ccagacgttc 240 aaaatcaggt tctaaatgaa ataaaagaag tccggggcga aggacagaaa attacgtata 300 aagagctgca agaaatgaag tatctggaaa tggtaatcaa ggaaagtatg agactgtacc 360 cttcggtgcc gttttattcc cgccaaacca ctgaagaagt actttatgaa gatggcaaag 420 tgattccaca aggaatcacg ttgatcatat cggcatacgc catccatcgg aatccacatg 480 tttatgagca accggacaag ttcatccctt cgcgattttt cgatttggag agcaaaccat 540 tcacttattt gcccttcagt gccggcccaa gaaactgtat agggcaaaaa ttcgccatgt 600 tattgatgaa gtttgccttg atcaacatgt tgtccaactt tgagattttg ccctcaaatc 660 caccctgtga aatggtttta tcagctgagt cggtgctgaa ggcccataat ggggtgaata 720 tccgcctgaa gtccaggact tagttgccta aggaatagtt aagcgatatt gtgacagtat 780 tagcaggttg tattagtctg gaggaaaaaa atagaataaa ttgttcttaa acatctaaaa 840 aaaaaaaaaa aaaa 854 // ID GT422656; SV 1; linear; mRNA; EST; INV; 787 BP. XX AC GT422656; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M04 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-787 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6b63e0eff8dfaf00b4587f69905b0b27. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 4. XX FH Key Location/Qualifiers FH FT source 1..787 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 787 BP; 264 A; 146 C; 171 G; 206 T; 0 other; ttttacattt gcatggacct gcctggtcat ggcaaatccg acccattgcc accaatactt 60 cccatccact ctgcggacta tctgctggca ataaaggtgg ttgtcgatta cttccaaagg 120 gacaaataca tttatatggg acacagttat gggggacaaa tgggtatgct gttctcgcgc 180 ttgtatcctc atctaatcga gaagctaatt ttactggata cattgcattt tgtgacgaaa 240 ccagctgatg cttttaaatc cgtaatgata acggcgattg aggaaacgct ggagttggac 300 agaaagacca aaacgagatt gccgcctcag tatacttatg aggaagcagt ccagaaagtt 360 atgcaagccc gacggagtcc gatatcagaa agagcagcga aaaatctact gaagcgatcg 420 ttgaagaaag tgggcgaaga taagtttgaa ttgactatag atcaacgact gaaatttgtg 480 ttaagcccac tgcatgataa tagatatgca attgaagtat taaaaagcgc tccattgcaa 540 tgtccagtcc ttattatttt aggtgaggac agtaaaatct tacggagttt tttactaccc 600 gttttagagc acttaaagcg gcaaaaatgc gtcaaaatag tgtacgtgaa aggagatcat 660 gatgtgcatc aagtgacccc cgagagagtg gcaccctttg tttgtgagtt tcttaattac 720 aacaaaagca aattgtaaca tcgaaatcag gaataaaaac ttttaaatga caaaaaaaaa 780 aaaaaaa 787 // ID GT422657; SV 1; linear; mRNA; EST; INV; 711 BP. XX AC GT422657; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-711 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2a566f3804199e65ae0cf77878968b98. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 5. XX FH Key Location/Qualifiers FH FT source 1..711 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 711 BP; 265 A; 155 C; 165 G; 126 T; 0 other; gaccaattga tggtgttgga aaacaaccgc aaaaacacaa actcgtccag tgacagcgcc 60 ggaggcccgc gaggtgaccg cagtgctcaa cgtagccaaa cgcgggcttt cagaggaaaa 120 gagagcattt tcaaacggcc ccagaacccc gcacctaaaa gctacatcaa caaaatacca 180 gacttcaaga aaaacccgca caaatggaca aggtattccc ttgaagacgt ccaggaagtt 240 acagaagaaa gcaatgccaa atccgcgatg gagttcctac gggagctggc aaaccggaaa 300 aagctggaga agggaatcgt tgagaagctg ggggaattgc cctcgaaaat cgttttcaac 360 aagcacattc ggctgacaga aacctcaggc accgctgaac ctgtgatgat aatggaaaaa 420 gctccgacaa gctttgaacc agcgcaaaaa ccctcgtata aaaacgcaaa gctagtgatg 480 cccgagtatg tgattggaca gaaagtggcc aaaaaggaaa aaagatccgg taaacccaaa 540 ccggtcaaag gaaaagagct caaactggat catttcatga ttgaaggaga ctagagattt 600 acccagttta agtcataagg cacatttagg gctagtgaaa tgcgataatc tccattatag 660 attcattggc tattaaactt aggtaagagc tataaaaaaa aaaaaaaaaa a 711 // ID GT422658; SV 1; linear; mRNA; EST; INV; 833 BP. XX AC GT422658; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M06 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-833 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f731a0b738f56ceefb6d6ced81659f2d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 6. XX FH Key Location/Qualifiers FH FT source 1..833 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 833 BP; 275 A; 145 C; 218 G; 195 T; 0 other; aatagtggaa agcaaaagcg cgaaatttcc cgttgggagg tatgtggttg gtcagtatgg 60 ctggcgatcg catacggtgg cgtctgaaaa tgcaccgcag gaattaggat tgcaaacatt 120 cctcttacct gatttacaag ggctgcctgt ttccttgggt ttaggggtgt tgggaatgcc 180 agggaatact gcgtattttg gacttttgga actgtgcaaa ccgaaagcag gagaaactgt 240 agtggtctca ggagctgcgg gagctgttgg aagcctcgta ggacaaattg ccaaaatcaa 300 aggctgcaca gcgatcggta tagcgggctc tgatgagaaa ggaaagtggc tcacccaaga 360 gctgggattc gatcacttta taaattacaa aacccaaaac gttgagaagg aattgaagaa 420 agctgcccca aaaggagttg actgttactt tgataatgtt ggaggagaaa ttagcacgac 480 agtaataaac caaatgaacc tatttggacg agtggcggtt tgcggagcaa tatccgctta 540 caatgcaaca ggcccaataa aagtgtccac ccccaatgcc agcattgtct ttcagcaact 600 gaaattggaa ggatttatcg ttagcacatg gaaccaacga tggatggaag gaattcatca 660 aaatctgcaa tggatccaac agggaaagct aaagtaccga gagacggtga ccgaaggctt 720 cgagaatata tttaaggctt ttactgaaat gttacaaggt ggtaatgtgg gcaaagccat 780 tgtaaaagtt tgagctaaaa aaatattaat tttgaagaaa aaaaaaaaaa aaa 833 // ID GT422659; SV 1; linear; mRNA; EST; INV; 884 BP. XX AC GT422659; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-884 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 86bbcb704fd6a18b1b4762e730bc8efa. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 7. XX FH Key Location/Qualifiers FH FT source 1..884 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 884 BP; 284 A; 181 C; 213 G; 206 T; 0 other; gggacccaga caatccgaaa gatgtgcata tcatgtccac gatcagcgct gctgcgactg 60 cagaagagta tcagaacata gtaaatgaga atggagcccg atacaaagag ggcgattctg 120 catgggaact gtacaattac tgcgcctcaa gagatgggaa tattggaaca gattgcagtc 180 cagtagtttg ggtggaaaac gaaaactttc agttagcagg ggtaactgtt cataatggag 240 ctactgatgc tcaggctgtt gccttaagaa caggagctga taagatccat ctaatgaata 300 accgattctt gggcctccaa gacactcttg gtctgggcat tgtcacccgc gacatcacta 360 aagtggaaag agttcttgtt catatgtgct atatcaaagg cgaaatagat tacgtatacg 420 gaggagccgt cgctgtcttt aacatggtta ctttcaacac aggttcactc aaagaaagcg 480 gctcgaaaat cgtgtttgct cccgacactc ccgcaggacg ttcctttggg ttcctagtga 540 ttaacagtaa cattactggc gatgctacct atataggttc caacaggatc agtctggcta 600 gatcatggga cagaggaatt aaaagcgccg acttgtacgt tccaggcgaa tccccgaatg 660 ggcaattggt gattagggac tcaaacatag acgatgtgat taatatagaa gctccctatg 720 cggctgcagc taccagcaaa cggccatttt caactgatat taaacaggat cgggatttgg 780 atgataacac ccataacaga ctgtgggagt acaataacag tggtcaagga agcccaaagt 840 aatatggaaa taaaacgtat aatgacccaa aaaaaaaaaa aaaa 884 // ID GT422660; SV 1; linear; mRNA; EST; INV; 834 BP. XX AC GT422660; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M08 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-834 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 7d5b568d6667090b0e9ee7f4afc6f5ef. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 8. XX FH Key Location/Qualifiers FH FT source 1..834 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 834 BP; 324 A; 135 C; 162 G; 213 T; 0 other; gaaaacatcc gaattagtac gaagagcatt aaaagatgta tatcaaatga aaaaaccaga 60 tgcattaatg tttgcaaaac gaaatgacat cacggtattt gaagatccca caccattgga 120 acgacattgt aaaaaaaacg agtgtgccca ctttttgctg ggtagccaca gcaagaaacg 180 tccaaacaat ttaactattg gtaggatgtt caattatgca ttattggaca tggtagaatt 240 acatatagaa tcatataaaa gcttgatgga tttcatcggg cctaaaatga cattaggaat 300 aaaatcgtgt ttgttattca atggacccca atgggacgag tcagacgacc tgaagatgtt 360 gagatcaatt ttcattgact ttttccatag agagcaaatt gatgcaatca gattgcaagg 420 cattgaacac acattgagct tcagtgttac tcctgccggc atcatttgcc tacgatccta 480 taaagtattg cttaaaaagt caggattaag tataccacga gtagaactcg aggaaattgg 540 tccccgaatc gatttttcat tgcgaagaac aaaaccagca tcgcctgatc taatgaaaga 600 agcttgtaaa aaagctagag aattaatacc gcataaaaag aggaatatta gcatggatgt 660 gtttggcaca acacatggac gaattcatgt tggaaaacaa gaaatcctta gaatccagac 720 tcgcaaattg aaaggtctga agaagacagg agaagaaaaa tgggcaaaac aaaaaaaact 780 taaagttttg gaaaaataaa gttttctttt tttttgcaaa aaaaaaaaaa aaaa 834 // ID GT422661; SV 1; linear; mRNA; EST; INV; 884 BP. XX AC GT422661; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M09 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-884 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f448cb44c8aa487d61d3d954fe87fc47. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 9. XX FH Key Location/Qualifiers FH FT source 1..884 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 884 BP; 305 A; 163 C; 222 G; 194 T; 0 other; agccgctgtg agagtcatgg aggatgatga agttttaatg caggatatgg atccgtgctg 60 aatttagatg gagaagtgaa aatggacgcc agtgtaatgg taggagctga cttatccgcg 120 ggagcagtaa ccgtggttaa ggatattgct catcccatca gtttagctag attggtgatg 180 gagagaactc ctcatgtact tttagcaggc gttggggcga acaggttcgc aaaagaacaa 240 gggatcgtta gactagctga gggcagttta gtttcaccgt atgccaaact aacacttgaa 300 gagttcagga agcaacagag cactggcacg gaaattaaaa atccgggcga ggtcggcact 360 gttggggctg ttgcaattga caaaaacgga aaattagctg ccgccacctc aaccggaggt 420 tattttggca aaatggttgg cagaagcagc gatacttctt taataggttg cggtacttac 480 gcagacgaca acatcggcgc tgtgtccaca acagggcatg gcgagtcgat agccaaatat 540 tgcttagcgc acgctataat aaaagccatg gagtatggac ataaatcggc aagcgatgca 600 acaaatgaaa ccctcaacca aatgacgttg aagctaaaac atactgcggg agctataact 660 ttatctaaaa ccggcgaagt aggcattgga tttacttctg aaggaatgtc atgggcttac 720 aggaaaaaga gcgaactgca ttatggaata gagaaggggc aacatgaaat caaacaagtc 780 ttatgaaaat tctactgcat aagtgtaaaa accagcaacc tacaacgcga attgtaaaaa 840 cctgcataaa ctaaatctaa aaaactaaaa aaaaaaaaaa aaaa 884 // ID GT422662; SV 1; linear; mRNA; EST; INV; 843 BP. XX AC GT422662; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M10 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-843 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ee6cdb61c53c7f3c6664894587d018ea. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 10. XX FH Key Location/Qualifiers FH FT source 1..843 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 843 BP; 259 A; 162 C; 167 G; 255 T; 0 other; gctgaacgaa catatttgga aaaaacttta tctagcactt ttgaaaatga tgaaccagtt 60 ccttgggcaa aaatattgac gtcaatccac ttctgggcta taattgttgg tcatgccggt 120 tactatttat ttgttatgct tttggttacc gaaatgccca cattcctagc aaaagttatg 180 aagtatgact tacagtccaa cggcacagtg tcagcaatac caaaaataat tagtacagta 240 tctggcatat tcatttccat tatctccgac tatattccaa ggcacggact gctttccgta 300 ttgaatacta gaagattttt tcacctaaca tggacgcttc taatgacgac cgccgtgatt 360 tgcaccactt atattccaga gcagttgcga atatgggcgg ttgttcttta ttgtattgta 420 actatagctg acgagtctat ttcaataaca ggagtctcag tcaatattta cgatctttct 480 ccaaaatacg caggaattat cacgggaata tcgcatagta tttcgcaagg tgtgtctgca 540 tttgcaccgt cacttgttga ctgggtttgt accgacttga gcgatgccag ccaatggagg 600 acagtgttta taataacttc ggtaatcgca ctgtcgagtg ctattttctt tgcgatattt 660 gccagcagcc agaggcaaaa ctggggaacc gaatcggaag aagacgctcg aatcgaagcg 720 gttataaaac gaaagggctc catttttagc atttattcac ttgctgcata aatagtgcta 780 cgataaatga aataaatgtt tctgatgcaa ataaatatgg ttaatgcaaa aaaaaaaaaa 840 aaa 843 // ID GT422663; SV 1; linear; mRNA; EST; INV; 869 BP. XX AC GT422663; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-869 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3142f9b0c172f9306b56aff1b0305cd9. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 11. XX FH Key Location/Qualifiers FH FT source 1..869 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 869 BP; 253 A; 184 C; 184 G; 248 T; 0 other; ccgcaatcag gcgcgacaaa gaatacgaga acattatcag gaagcaagca atggcagcca 60 gagctgaagc ggaacgtgac gggatccaag tgccactcac actcgaagat tactgcataa 120 aaactcaagt taaacccagc aactccgagt ctcaagtaga aatgacggat ttctacgacg 180 atgactacga cgaggcctac gacgatctgt cagacaattc ggaatatggc gatgaagacg 240 acaatgacag cggtaacggc gaatcgtgat aacgcatgca catccagccg attgccacat 300 tctatgtaca tttaaattca atacaagcca ctcttttcct gatttcattc taaccgtgct 360 tcgatagttt ttcgggtgtc cgaaaacgaa tcgattattt ctactggctg gcggctgatg 420 ttttcagtaa acctgttcag gtcgttagca aataagtact gtaagcgttg tgctctcgat 480 ctcaatttga gttggtgatt tcaagatgac ggcgatgtgt gttaacgctc ttagattagg 540 ttcaaagttc gcctgtctgt atatagttgg agcagtctaa aaccctccct ctaaccgcta 600 gtttatctgt gaaactccgt tctcccccaa tggcgcaact tcattcatcg tccagcagtt 660 tcttccgatg ggcaagagaa tatgttttat gtaaatattc gtctttttgt cgtcactcct 720 ctaccagttg gaatgaagaa gcgggttatt ttttgacgca ttgcaaataa atcgttcgtg 780 tgtgtagttg atttatgtta cgccactttt ggaagattaa ctttagtttg cattattgaa 840 taaatgttta ccaaaaaaaa aaaaaaaaa 869 // ID GT422664; SV 1; linear; mRNA; EST; INV; 812 BP. XX AC GT422664; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-812 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4a573974d684de2e8d886dc930d6dc25. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 12. XX FH Key Location/Qualifiers FH FT source 1..812 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 812 BP; 240 A; 183 C; 223 G; 166 T; 0 other; ctgaggacag caagctgctg ctcgccaact accacaccaa gaaaaagtac cggcaaactg 60 tgatgttcac agcgacgatg ccgccggcag ttgaaagatt ggccagaacc tacctgagaa 120 gaccggcggt ggtgtatatc ggctccattg gcaagccggt ggagcgcgtg gagcaaatcg 180 tgcatattgt gggcgaaaac gacaagcggc gcaagctgat ggagtatcta tccaaaggcg 240 tggatcctcc aattattatc ttcgtcaacc agaagaaagg cgccgatgtt ctggctaaag 300 gtttggagaa gttgggatac aacgcgtgca cgctccacgg aggcaaagga caagagcaga 360 gagaatatgc cttggccagc ttgaagtcgg gagctaagga tattttggtg gctaccgatg 420 tagccggaag aggcatcgac attaaggacg tttccatggt catcaactac gacatggcga 480 agaccattga agattacact catcgtatcg gtcgtactgg acgtgcggga aagactggaa 540 tcgccgtcag tttctgcacg aacgacgact cggccttatt ttacgacctg aagcagatgc 600 tgttgtcgtc gccagtctcc tcctgtccgc cggagctgat gaaccactcg gagtgtcaga 660 ataaacccaa tcaacccaaa aggcgacggg atgagatgat tttcgcgtaa accaaggcgg 720 gcaactggat tgattgatac acgttcttgg actgtacaaa gcatctattg atagtaaaaa 780 atgtggtggt aaacaaaaaa aaaaaaaaaa aa 812 // ID GT422665; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422665; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3962074679d7fcf39df56bf3991b24da. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 13. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 250 A; 161 C; 156 G; 249 T; 0 other; ataaatccga gttgactgac gtgtctgcaa gctaatttgc cataagtttg aaactaaaat 60 gaattcggtg ctccaaaggg gcaactccat tctagcctat gcgcttagcg cgctggcctt 120 cctgacattt tgctgttttg cctccacagt ctttctgaac tattccacaa atgcagatat 180 caaaactgtg aaagttttgg ttaagaacgt gcctgatttc agtgcctcgc gggatgtaaa 240 cgacttgggg ttcctgactt tcaacctcaa aactgacctt actgatatat tcaactggaa 300 tgttaagcag ctctttatgt atcttaccgc tgagtatgtg acgaaaaaca atcagcttaa 360 tcaggtagtt ttatgggaca agatcatcct acgaggagaa aatgcagtgt tagacttcaa 420 gaatatgaat accaaatact atttctggga cgatggagat gggctaaggg gaaataagaa 480 catcacactg acactgtcct ggaacatcat cccgaatgcc ggtctattgc cgaacattta 540 cgccagcggc atgcattcgt tcaagttccc tgtagagtac acgtcgtcca aaatgtaatc 600 cattatcaag gttatttttt taatttccaa cttgttctgg ccgccactca aacaaaaact 660 attttgtgta tatttactta tttaaattgc aaagcatttt gtgtgttgtg cttgaaaaat 720 acgttttttt ataaagttag tcttttattc atcaagtgca caagggtttt cctacataca 780 taacatacag tgagtagtga aaaaaaaaaa aaaaaa 816 // ID GT422666; SV 1; linear; mRNA; EST; INV; 824 BP. XX AC GT422666; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-824 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 05ba1e94357d147f89fbf12efb5718ff. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 14. XX FH Key Location/Qualifiers FH FT source 1..824 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 824 BP; 302 A; 121 C; 127 G; 274 T; 0 other; agcaataaca ataaagtaca catacagggt gaaagcgata aaactgcaga aaatattgtg 60 tctgttagtt acgtatcggc tgcaacccat gcggctaatc aaacgcacca aaactcctgc 120 acttttgaac tcaggcattg agggccttta accggttccg tttgccagaa attatcatcg 180 gaaaactaat gcaatctctt tgtgtaaaat gaagaatttg ggagaaatta attaaaactg 240 aatctgtcga tttttaagat tctaaaaaac tgccgcatga attgccgtta ttttttgcat 300 attaatattt gatcatgaaa atcacaaact tgttaaaagc atgtgctatt tttgcaaatt 360 gtatataaac gcactaattt ggaaacgatt cgaatagtca aaacactatt ttccagttaa 420 tttcaagcac ttcaaatgat atttgaagcg cataaataac tttttttcca taaaatttta 480 agcgggtgtt aagcagtttt tacacaccct gcataaacat tccacttttt ctttaaatat 540 tacccgcagt agttttaggt gttaagcaag tgcgaaattg ttcctgtaca tttaagagaa 600 aattgtcact taggctgtgt ttatattatg acgttgccaa tatatttatt attaggtttt 660 atttattttt atacattagt gttaataaat cgagcaattt ttgttacgta aactcgatgt 720 tatgttatgc attaagccag taaaatataa aatgttttta tataaaggaa gcaataaatt 780 tttggtaata gaaaaaaaaa aaaaacaaaa aaaaaaaaaa aaaa 824 // ID GT422667; SV 1; linear; mRNA; EST; INV; 832 BP. XX AC GT422667; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M15 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-832 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 670b0e1148109355f5ee5c124647b939. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 15. XX FH Key Location/Qualifiers FH FT source 1..832 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 832 BP; 280 A; 145 C; 160 G; 247 T; 0 other; ggggggaaat caaacaaaca cctcttgcag attaattcct tatttcctca tatttaagaa 60 caaaagtaca ataattccac atatttataa tgctagtttt agttttggga gatttgcaca 120 ttccacatag atgtagtagt ttgccagcaa aattcaaaaa actcctggta ccaggaagga 180 tacagcatat tttgtgtaca ggcaatttgt gcacaaagga atcctacgac tatttaaaaa 240 cgctggccgt agacgttcat gttgtcagag gggactttga tgataacatc aattatcccg 300 aacagaaagt ggtgactgtt gggcagttcc gaatcggact gtcgcatggg catcaggtgg 360 tgccgtgggg cgatcctgag gcattagctc ttgtgcagcg acagttggat gtcgatattt 420 tgatatcagg tcatactcat aagttcgaag cttatgaaca tgaaaacaaa ttttacatca 480 atccaggcag tgccactggc agttacaatg ctttagatat gtctatcaca ccctcttttg 540 tccttatgga catacaaaac actacagttg taacgtatgt ttaccagctt gtaggggacg 600 acgtaaaggt agaaagaatt gagtataaaa aaaattgatt tggccaccca agtccctttt 660 caaacgcgaa tatttcttat tattgataat tgaagattaa aggtataatt tattaggtac 720 actttatacg aggaaaatta tttattaaag gacactccat taatgttata caccgcttat 780 gttacaccaa aagaataaat ttttatttac agcttaaaaa aaaaaaaaaa aa 832 // ID GT422668; SV 1; linear; mRNA; EST; INV; 857 BP. XX AC GT422668; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-857 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4e68a2e1b533c95a89e7826f30f3e4c7. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 16. XX FH Key Location/Qualifiers FH FT source 1..857 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 857 BP; 307 A; 117 C; 160 G; 273 T; 0 other; atgactttta atggattgaa atgggctgga acacttacca aatatgttac ttcttaccgt 60 cactatctca ggaacagctc aagactgttt tctgtaaatc ggccagaaag cgaaccagtg 120 ttaagcaaaa aagaagttca agtcacgttt ataaaagcaa acggtgagaa gatccaaacg 180 aagggcaaaa taggagattc gctcttagat gttgtagtca ataacaacat agatttagat 240 ggattcggag cttgtgaagg aactcttact tgctcaactt gtcacttaat attgtctaag 300 agcacatacg attcattacc aaacaaacct tttgatgagg aattggacat gttggattta 360 gcctatgatt tgtgtgatac atctcgtttg ggttgtcaaa tagttctaac agaggagtta 420 gctgggctgg aagtaaatgt acctgctaca ataaacgacg ctcgcagtta acattagtag 480 gacgagacaa tagatctata taaattaaag gaaacattta atgcaatttg ttagaagtta 540 acgttatccc ataaagttag agatttaaca ttaaaaatat ttttgtattt agtattatcg 600 aactagtata cctactaatt ggaatgtatg tttccctcac aaaatgagac attgatttag 660 tttgtatata tgtacttaga cgattttata tatggtcaat tatgataatg acagttgcat 720 gaatagtaaa ttgttaatcg ttgattaatc gacacagtgt ttttttaatg tttttgtcat 780 ttttagtgga aaacttaaac atttgaagtt tgtgaaaaaa taaacgaaca tatttgagga 840 aaaaaaaaaa aaaaaaa 857 // ID GT422669; SV 1; linear; mRNA; EST; INV; 722 BP. XX AC GT422669; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M17 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M17 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-722 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 84d0fa3aad9154d012dca6e8bc1ff2a2. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 17. XX FH Key Location/Qualifiers FH FT source 1..722 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M17" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 722 BP; 230 A; 135 C; 145 G; 212 T; 0 other; ggggatctga cagttatcct tcgtatctgt tgtagatatt tttgagtttt tagaggaata 60 tcgtcggata atctgattta gaaaaatatg gctgcagtgc tgaaccacgc attgaggaga 120 ttcttccaaa catctgccca gcgggcagtc caaatcgagg gacattcagc agtgtctggc 180 atccatgagg ggggctacaa accatggaag aaacttacgt tctttgtcgc attcccttcg 240 attattttgt gtgctgtaaa ctgctacatg gttcaccaag atcatgccaa acatccacat 300 gagaagaagt tcgtcaaata cgaatatctg gcaatacgta gcaagaggtt cccttggggc 360 gatggcaacc atagttttgt ccataacccc aaggtaaacg ccctccctga cggctatgaa 420 gcataagcaa cattcgtata atggtagttg tttagtcagt ccagttattg ttcaattaat 480 tccttattta tattaaacta gctcacaaat atgttcagta gagttattca gtccgccatt 540 ggattggggt atcaataagg agtattttaa atcaacaata gttttattgc aatgtacgaa 600 taatcgagaa aaaacactag tcttattagt aatgactggc gctttatttc agtcagttag 660 gcaaaaaaat gcagtttact ctaacaataa aatcaactgt tgcatcaaaa aaaaaaaaaa 720 aa 722 // ID GT422670; SV 1; linear; mRNA; EST; INV; 874 BP. XX AC GT422670; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-874 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 77643ea3d0d3c508565f0ab9b38eeeab. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 18. XX FH Key Location/Qualifiers FH FT source 1..874 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 874 BP; 284 A; 169 C; 183 G; 238 T; 0 other; aactgatgat gataaagtta ccgggctccg gtaacggaaa aggaatttta tcggaaagcg 60 acattatcgc tcagtgcttc gtcttcttca ttgctggctt tgaaacttct tccagtacca 120 tgacttttgc catgctggaa ctcgctcaac atcaggatat tcaagacaaa ttgagaaagg 180 aaattgtgga ggttttggcg aaacatgatg gaaaaatcac ttatgaagcc gtaatggaaa 240 tgccctattt ggacaaagtc atctctgagg acttaagaaa gttcccacct ctacccatca 300 ttccacgaag atgcactaaa acgtacaaag tgcccggaac tgatttggta ttggagcgcg 360 gcacagatgt gcaaattcca gtttgggcca tccagaacga tcctgaatat tacgagaatc 420 cagaggtatt cgatccagag cgattctcag aagaaaacaa agccggcagg ccggaatacg 480 ctttctttcc tttcggtgct ggacctcgcg tttgcattgg cttgagactt ggcaaacttg 540 aaactaaagt cggactaatt acgctgcaga gaaaattcaa gttcaccatc aacaagaaaa 600 cgccattccc cattcagatg gagaaacgag gatttatcag cactatccaa ggagaccttt 660 gggtcgatgc ggttagaata taaggcatgt gaactaagta ttcctgcggc gttattatta 720 atatttttaa tatttattcc cggttgtgaa catattctta atttgttagg caattattta 780 aactgataag acaagagttg cattattaga gtaataaatg tttttaaata aatattcctt 840 aacagcatgc cagatggaaa aaaaaaaaaa aaaa 874 // ID GT422671; SV 1; linear; mRNA; EST; INV; 851 BP. XX AC GT422671; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-851 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; faf31088659f6e5c01744bc8a13020ce. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 19. XX FH Key Location/Qualifiers FH FT source 1..851 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 851 BP; 273 A; 121 C; 156 G; 301 T; 0 other; gccatgactg tgatattaag ggaaatatag cagctaatat tggtttcgaa aatgaagcag 60 ttgatgcgag taaatttttt tggtgggtgg taggtatttt actattaaca agttcattac 120 tactgtcggc tcgaatgggg atctaccagg aaacgctcta taagaagtat ggaaagcacc 180 ctgaagaagc cttatattat acgcatttat attcacttcc tggatttctg ctatattccg 240 gcagcatatg gaaccattcg atagtagcat ccaacagtga tccgtaccag ataccattta 300 ctagcatcgt aatatcagta atttggttat atttaatttt aaatgtgctg actcaatatc 360 tttgcattag ctctgtttat gtattgactg ctgaatgtac ctccttaact gtcactttag 420 taattaccct acggaagttt ttgtcattgg ttttcagtat cgtttatttt caaaatccat 480 tcactacagc gcattggatt ggaacagctt tagtatttgg gggtactctt atatttactg 540 aggttccgaa aagaatatta gaaagcagaa aacaagttaa agaaagtatt aagaaagcta 600 attgatagat tatttttgtt ggtttagtaa atatgttcag tcaaaggtct tgttttatgc 660 atagacgtta tatgttattt gttcaaaagg gcatttttag ttaatacctc atagttcttt 720 agtgattagt tagttagatt aataataatt atagcagatt aatatattcc tggtgtatac 780 gaccgactca gtttcaggtt ttattatagc aaattaaaat gaattctgaa gtcaaaaaaa 840 aaaaaaaaaa a 851 // ID GT422672; SV 1; linear; mRNA; EST; INV; 582 BP. XX AC GT422672; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-582 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; bfaf6592c2d49c815ed6af3920ee0961. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 20. XX FH Key Location/Qualifiers FH FT source 1..582 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 582 BP; 191 A; 113 C; 135 G; 143 T; 0 other; ggggaacatt cggttggctc tcctgtaaac tgtgataacc tctttctgag agtagtgttt 60 atcaaaatgt cgctggtgat tccagagaag ttccaacata ttctccgtat cctcggtacg 120 aatatcgatg gaaagcggaa agttatgttc gccttaacgg ccatcaaagg agtgggtaga 180 cggtactcca acattgtgct caaaaaggct gatgtggatt tgaacaagag ggccggagaa 240 tgctctgagg aggaggtcga aaagatcatt accatcatgt ccaatccacg acaatacaaa 300 attccagact ggtttttgaa taggcaaaaa gacattgtcg atggaaagta cagccaactc 360 acatcatctt ccctcgactc gaaacttcgt gaagatttgg aacgaatgaa gaaaattcgt 420 gcccacagag gtatgagaca ctattggggt cttcgcgtga gaggccaaca cacaaaaacc 480 actggacgtc gtggacgtac tgttggtgta tccaagaaga agtagacgca ctcgtgttta 540 ttttttaaaa ttaaattcta aagtcaaaaa aaaaaaaaaa aa 582 // ID GT422673; SV 1; linear; mRNA; EST; INV; 902 BP. XX AC GT422673; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-902 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9f833e32c548db5b4123793aa880ea2c. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 21. XX FH Key Location/Qualifiers FH FT source 1..902 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 902 BP; 283 A; 168 C; 151 G; 300 T; 0 other; atacagttaa catagtagca tcaatgatgc tattttcaag atttcttcat aaatacatga 60 acacttgttt agctggagaa ggatctgtgg gaattttggg aaatttattg aatataattt 120 ccatatcaat ctttaaagat gacgtcacaa actcaacttt gctctatttt attattggca 180 ctgccattct atgctgcacg tgggtcctaa ctttctttac gatgaagtca gaatttttca 240 tccacagtat tcagtcgcta ccagaagatg tcaacaaaac gatgcctggt aaaaagcaaa 300 tcatagaagt attctcaaaa atcaaatttg ctgtagtttt atttgctata tttgttacta 360 gctacgcagc tacacataca gctgtaactg cactcgtagt ttcagagcag acggatacca 420 catggtctac taattacttc agtcccgtga tgacgttctt cctatcggac gtgtgcatgc 480 tagcaggccg tttaatggca tcagccataa ccatagatat cccggaaggt ccatttttat 540 tagtcagcat agttagaacg cttgttctag tacccctcat atggctttgt aatgctcaac 600 ctagaaatca tctaccagtt ctatttcccc atgattatca atacggtatt atactggcga 660 catttatgct taccagtggc cttatattaa acatgtccat actggtggtt ccaaagaaag 720 taagcaaaga agatgctgat gtagcttata cattgattgg tttaataata aacacgctcc 780 aagcactaac atcacctata ggtttagtta ttgttaatgt tttgtagtta atttctgttg 840 cttttgttgt ttttatattt atactactat taaatagagg aaacaaaaaa aaaaaaaaaa 900 aa 902 // ID GT422674; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422674; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 83f611c3e2995645bbf2728f7c07ed93. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 22. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 183 A; 219 C; 198 G; 216 T; 0 other; aatatcaaat ccctggatct tttcgccgag cccattggcc cagcccctcc tcaaccaggt 60 ggcctcagag tgattggagg aacagttgct gcccgtggcg cttttcccta tcaagctgcc 120 ttgattgtga accttctcag tttctgcggt gcctccatca tcagtggcga ttggatcttg 180 actgctgctc actgtgttga ctcagctttg gtggttcaag tggtggtggg cgcacataac 240 cccagaacca ctgttgatga acccactcag gtccgattga gcgccaacgg ccgtgttatt 300 cactccgctt gggacagggc gaatcttcaa aacgacatcg ctcttctgag agtggccggt 360 atcccagtag gatctcctgg aatttcttcc atttccctag ccccagccac ttccggaact 420 ttcgctggat caactggtat cctcactgga tggggaagaa ccagcgatgc cagcaatgct 480 gtatccaacg aactcagaac agttcaactc cccattctga ccaacgcagt ttgccagagc 540 tctttcggca gcatcgtcac tgctcagcac atctgcacct caggggccgg aactcgagga 600 gcctgcaatg gcgactctgg tggcccattg gtggttagcg gtgttgaagt tggtgttgtt 660 tcatttggag ctagtcaatg cgaagcagga catccctcag tgtttgccag agtttcctac 720 ttcaggaact ggatctcatc caactccggc atctaatgat tttgagcgag cgatttgatt 780 aaattttgtt tttttgcgaa aaaaaaaaaa aaaaaa 816 // ID GT422675; SV 1; linear; mRNA; EST; INV; 812 BP. XX AC GT422675; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-812 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 91a46f9716c928db4cf30625b7174314. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 23. XX FH Key Location/Qualifiers FH FT source 1..812 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 812 BP; 222 A; 179 C; 224 G; 187 T; 0 other; aaggcgagga cttccagaac ctattccagg ctgggtattt cgtagggggt tgggccccag 60 gaggaaattt tactgtggaa agaggcttca gggagttgga tcatgggcac gacttctacg 120 cctcccagac gtttcaggcg cccgatggga ggagactgga aataggctgg atgggaatgt 180 gggagtcgca attccctgaa aactcctcag gctgggcagg gatgctgtcc ctgccccgag 240 agctcacgct ctccgatcag ggagacttgg aggtgcggcc cttgcgggaa attcaaaacc 300 tacgcactga gcggctggag gtttctcaaa ctctccacat agcccctgga ggaggacttg 360 agatactcca aaacatctcc catagtgaag ttgctttgga cttcaacctg ctcaactcca 420 gcagcaattc attcgcagtg cagctgacca gggagagttt tgaaagcgac ggaggcgctc 480 aggtgtatgt agatcgcaac tgcagcagga ttttcctgga acgccactat cctgcataca 540 atatagagag aagcagcagg agtgtagctg taaacctaac tgggaatttg tttttggata 600 tattcatcga tggctcttca atggaagtgt tcgtgaacga tggccaagcg gtgatgagca 660 gcaggatcta cccagacgat cagctcagaa cggttctgat cactgctcag aatggatcag 720 tggcagtcga tagacttaca gtttgggacc tgtagttaac aattgtaaat gctttttaaa 780 cagttatgaa attcaaaaaa aaaaaaaaaa aa 812 // ID GT422676; SV 1; linear; mRNA; EST; INV; 885 BP. XX AC GT422676; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_M24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_M24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-885 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5eade438fc529d99c071a29d4459d3dd. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: M column: 24. XX FH Key Location/Qualifiers FH FT source 1..885 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_M24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 885 BP; 316 A; 131 C; 152 G; 286 T; 0 other; tgaaatggag gaggaatcag atcaatgtgc gaaaaggttc ttagataatg agattgaact 60 tgatagtttt ctggacgagt ttcaaacaaa acgaaaattg atgcataccc gattagtcaa 120 agcggacaaa ttggctaaaa tattgcaaag aggtgatcca tccatagggc tgggaatgcc 180 caattatata aatgcgcctc cagttaatat aaatagggga ttctttccgg gggtagtttc 240 tccaggttct ccgccagtgg gtggcgtgcc ttatccaacc gggcctctga atatgccttt 300 acctcctctt ggaatgaact ttcctcagaa tcattattga aaaattctat attaaacaac 360 ttttcatttt tatcgcgcaa aataaagtgt ctaaaaaact gattaaaggg actagagata 420 agtcgttgct tatgtactgt agagcaatgt aaatcgaata gttgtgaaga aaatttgatc 480 agcatatttt ggacactgcg cgtttgttat tgaatgcata tttttatgaa agagatttac 540 ctactttata gaatcactat gaataaattg tagataatat tcaattaaca ttattcactt 600 agaagccatg ttaggatata tgttccgtat taaaaagtaa aatagatttt aatgttggtt 660 gcataacttc aaacaataat atatgcatat gtaacatttg tgggtatttt gtaaattctt 720 gcctgtgatt tcactaaaat cctattttca cttttctaca tgaatcattt taaatcggca 780 acttgaaaat aagaactaat tagcaattaa atcaaattag tcctaaacaa atatttattt 840 acagaattaa tatagtacat tacacagtaa aaaaaaaaaa aaaaa 885 // ID GT422677; SV 1; linear; mRNA; EST; INV; 856 BP. XX AC GT422677; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-856 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 135c441f614cd1a3d0650daee8b78478. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 1. XX FH Key Location/Qualifiers FH FT source 1..856 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 856 BP; 320 A; 125 C; 171 G; 240 T; 0 other; ttatcaatgt tgttttagat taaatttatg tgtgtttgtg tagaatgtgc ggcaaacaac 60 agggctacta catcttttgc agaaaattag tgaaaaaaca caatttttaa acatgtttgg 120 ttcggggaat caagaacagg atgaaggcga tggtaaacag aaattgaaag atgtggatcc 180 acctaaggac tctgaagaaa tcaaagacat gatttatgat ttgttaagga atattaagga 240 cccagaaaag cccaacactt tagaagaact tggcgttata tacgaagacg gaatttttat 300 aaaagcaccg actggaggcg gcgttaatgt aatccgagtg gaatttaacc ccacagtacc 360 ccattgctcg ctggcaactt taattggact gtgcataagg gtgaagctgc aaagagatat 420 gccttttcct ataaaactga atatttttat caaggagggc gcccacacaa cagagcatga 480 gattaacaaa cagattaatg acaaagagcg aatagctgca gccatggaaa acccaaacct 540 aagagaaatc gttgaaactt gtatcaaaga tgaagactga aaatgtctgt cgaaattcgt 600 attttatgat aaattaaaga tcggctaatt ttttaaatgc gctaacttac gtcatatatg 660 cgaactcaca ttgaaatcta tgtgagtttg gcaaagatga aggtcggtac tattggtcac 720 gatttatcgt ctgaaaaatt gattgaaaat agattaagta gacccataag ttgtaataca 780 attatatttg ctttcgtttt ctatatgaga aaaataaagt ttaaaatatt gttatagcaa 840 aaaaaaaaaa aaaaaa 856 // ID GT422678; SV 1; linear; mRNA; EST; INV; 830 BP. XX AC GT422678; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-830 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; dfd1462e214b6f4fd46bfe1cbc70fcbf. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 3. XX FH Key Location/Qualifiers FH FT source 1..830 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 830 BP; 266 A; 168 C; 188 G; 208 T; 0 other; acggccctcc tggctgtggg aagaccatgt tggctaaagc cgttgctcat catacaactg 60 ctacatttat cagagtggta ggctctgaat tcgtccaaaa atatctgggc gaaggtccca 120 gaatggtaag agacgtgttc aggctggcga aagagaactc acccgccatc attttcattg 180 acgaaatcga tgcgattgcc accaaacggt ttgatgccca aactggggcc gatcgagaag 240 tccagaggat cctgctagag ctacttaatc aaatggatgg gtttgatcaa acgaccaacg 300 tcaaagtcat tatggctact aatcgggcgg acactcttga ccctgcccta ctaagaccag 360 ggcgtttgga cagaaaaatc gagtttcctc tgccggacag aaggcaaaag aggctggttt 420 tcagcacaat cacatcgaaa atgaacttat cagaagaggt cgatttggag gattacgtag 480 ccaggccaga caaaatctct ggagccgata tcaacgctat ttgtcaagaa gcaggaatgc 540 acgcggtgcg tgaaaatcgg tacatagtct tgcccaagga ctttgaaaag gggtacaaga 600 acaacattaa gaaggacgag agcgaacatg aattttacaa ataaagtttt ctctgttatg 660 tgaatggaac taaacatgcc tgcagttcgg atttgttgca caactatttt cgtattgccg 720 gataattgtc aataaattta gataaattag gtgttcaaca atattttaca cccaattttg 780 tttttgaaca ttaaaaatgt cctttttctt cgaaaaaaaa aaaaaaaaaa 830 // ID GT422679; SV 1; linear; mRNA; EST; INV; 617 BP. XX AC GT422679; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N04 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-617 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f9dde30af685f7d62da9645919c4b754. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 4. XX FH Key Location/Qualifiers FH FT source 1..617 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 617 BP; 204 A; 120 C; 135 G; 158 T; 0 other; gaaggaaaac tcataaccaa gtatcgcaaa atgcctttat ttgacataga tattcctgga 60 gggataactt tcaaagaatc cgatgtgtta gctgctggaa acaacttagc ttccttcgat 120 ttagaaggta ccaaaatagg cctgggaatt tgctacgact tgcggtttga ggaactagca 180 aaactcttcc gattgcaggg agtggaaatt ttaatatatc cggcagcctt caatttgacc 240 cccggccctt tgcccttcga gttgctgcaa agaggcagag ctatagataa ccaagtgttt 300 gtcctggccc ttagcccggc cagaggcccc cagggctata ttgcttgggg gccctctcaa 360 atcccaaacc cctggggcaa agttattggg caaaccaaag aaggggaaga gattgtgaat 420 gttgatcttg acctaactga atgtgataaa gttcgacaac aaataccaat tttcagtcag 480 cggcggaatg atatatacga tacttttaaa gtaggaatta aatgataggt agaaataaga 540 taaaaacccc gtaaaccgaa ttaataggtg ttttatggag aatatacttg tatcccttta 600 aaaaaaaaaa aaaaaaa 617 // ID GT422680; SV 1; linear; mRNA; EST; INV; 820 BP. XX AC GT422680; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-820 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 1ccb8087ce6950f805de6301902a3916. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 5. XX FH Key Location/Qualifiers FH FT source 1..820 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 820 BP; 269 A; 154 C; 203 G; 194 T; 0 other; aggggtggac gccagggctg agggcgtcga aaagaagatc aatgtgttgg aaaacgacct 60 gcggaagttc agggagcaaa tgtccaagat gcgagaagga ccagccaaga acgcagtcaa 120 agctaaagca ttgcgaattc ttaagcagaa gaaattgtat gagaatcaac tggataatct 180 caggggccag tcctttaaca tggaacaagc gaactttgct catcaaacgt tgaaagacac 240 ccaaactact gtaattgcca tgaaagacgg catgaaagcc atgaagcggg actttaagaa 300 gatcaatatt gatgatattg atgatgtcca agatgaactg gctgatatgc tggatcaagc 360 tgacgaagtt caagaggccc tgggcaggag ctacaacact ccagagttgg acgatgatga 420 attggctgca gaactcgatg ctttaggcga tgaattagct ttggatgacg atcagagcta 480 cctggatgat attgggaaat tgccagacgc tcccataacc aagcccgatg ccattgccaa 540 cagaacaaac aaggatggcg tgcccgtgga tgaattcggc ttaccacaac tgcccagcaa 600 tcattgagtt aagagagcag gtgtatatta agcctcattc aaagtacatc taggcatgtg 660 gttaaagccc gggatttaac accgagtcgt ttttatattt tattgtggtt aattggcact 720 gaactagttg ctggatcagc aggtttattt ggtttccacc cgcactttgt gttaataata 780 tatatagtaa cgatttttat agaaaaaaaa aaaaaaaaaa 820 // ID GT422681; SV 1; linear; mRNA; EST; INV; 783 BP. XX AC GT422681; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-783 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6aa176547eb5563f9113bed060c87255. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 7. XX FH Key Location/Qualifiers FH FT source 1..783 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 783 BP; 261 A; 151 C; 159 G; 212 T; 0 other; gggggagcca tttgggttca taaactttga tcgcgaccaa ttttctgaaa aattgtaata 60 aatcatctaa caattcaaag ctctgttaaa atggcttgcc gaaaagtact atccatcagg 120 caattcagta ctggccaagt aaccagtcag ttggtcaagc ctccagtaca aatatttggg 180 attgaagggc ggtatgcaac agccctgtat tctgcagcca ccaagaagaa gaccctggac 240 agtgttgaaa aggatctgat tgggctcgag acatctttga aaactgatcc taaattcaag 300 agctttatac tgaacccaac aattaaaagg tcattgaaat ctgaagccct aaaagcaata 360 tctgcaaaaa tttcgcttaa acccgaatct gccaatttgc ttcagctttt ggccgaaaac 420 ggtcgtctca taaagttgga tcgagttatt accgcattta aaactatcat ggcagcccat 480 agaggagagg tagtttgtga ggtgactagc gccagagaac tggacgccga acagaagacc 540 aaacttcaag gagtgttgaa atctttcttg aaatctggtc aatccatttt gctaactact 600 aaagttgatc cttccattat tggaggtttg cttgtttcta ttggtgatag atatgtagac 660 atgtctgttg ccagtaagat taagaagtat tctgaaatta tcagcgcccc cgtttaaatc 720 gaccaacggt gtaagcacga attagaacat aatgcaatat ctaattaaaa aaaaaaaaaa 780 aaa 783 // ID GT422682; SV 1; linear; mRNA; EST; INV; 841 BP. XX AC GT422682; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N08 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-841 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c5812c0bbe3ad77b6435f41378f099eb. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 8. XX FH Key Location/Qualifiers FH FT source 1..841 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 841 BP; 279 A; 152 C; 186 G; 224 T; 0 other; aagtatcaat cataaatgcc ccgatggggt cacaatacac atgtacttta ttcatcagcc 60 taaagaccga caacgacttg ctcctacttt tgttttcttt catggcaatg ctggcaatat 120 gggtcacaga ttgcaaaact gtgcaggctt atatcacaat ctgcactgta acatcctgct 180 ggtagaatac aggggatatg ggctagcaga gggaagccca tcagaagaag ggctgtatat 240 ggatgccaga gccagtttgg attacttatt tagcaggaat gatattaatc attcggaagt 300 tgtggttttt ggaaggtctt taggaggtgc tgttgcaata gacctagcag taagagagtt 360 ttacagccat aaaatttggt gcctaatagt ggaaaacaca tttaccagtg tgcccgatat 420 ggccaaagtt ttgctaggtt ggaaaatttt gcaatatctt ccgatatttt tttacaaaaa 480 taagttccag tcttatcaga aagtaaagca actgagaacg cccacattgt tcatttctgg 540 ccaatcagac actttggtgc caccgaaaat gatgcacgaa ttgtacgaaa ggagcaacag 600 tgtacgaaaa caactgttcc aactaccagg aggaacgcac aacgaaacct ggcaattgcc 660 aggatattat cacgctatcg cgctgtttct gcagaattac agaatatcag ctctaatgca 720 acaagctagt atagatatag atgcgaaatc ggacagtagt agtggaagtt tgtggagtag 780 tgtccaggat atatgagaaa aatggtcagt taaatcattt gaagaaaaaa aaaaaaaaaa 840 a 841 // ID GT422683; SV 1; linear; mRNA; EST; INV; 887 BP. XX AC GT422683; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-887 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3e7682dbea87ffdc9b145f2dfbe34edb. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 11. XX FH Key Location/Qualifiers FH FT source 1..887 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 887 BP; 318 A; 162 C; 156 G; 251 T; 0 other; aacatgcatt gcatgtggca acagaacgtt atacttgaaa tcaatgcctt ctccatattg 60 ctccaaatgt ttcagtcagc caattccagt tgcttataaa aaatcatcat ttacgcaacc 120 tggaaaatgc gtttgcattc tttgtaaatc ccaaatcagc gttacggagc ggcgacgcca 180 tcaacacgaa tgcaaatacg gaaaaccaca agtgtttgtc gaaaaactgg atatacacaa 240 gatagtggat attggaaatt cgactaaaaa gtgttcgagt tctagtggta gcgatatgga 300 tggttcacca cagaaacatc ctgcaagttc aagcgacgaa atacgtccac gaaaacgaca 360 caaattttca agtccgaaga aaccacctgt ccaaaatgat ttaatagcgg ataaaccaat 420 atcctttgac ggtacttacc agtgtcggtt atgcgaatat aaaaacacga atcgcaaagt 480 tttccatgag catataacaa ctcatagaaa tatttccaca gcttaccaat gtatggagtg 540 cggcgaatgc ttcgtggtca aaccttcatt aattaaacat ttagtacatt tccataatat 600 tctaaatgat gacaaatatt tccaacagaa tgattgtttc gacaaatgtg ctgtaacgga 660 attagctaaa gttgttaagg ctcctcattt ggcaaataat gttaaagata atcaatgtca 720 agtgtgcatg gctcagttta acactgaaga agaacataac aagcatttta gatctcatgg 780 catggctttc ctaatgaata agtcgattta aataattgtt gataaatatt ttttaagaac 840 ttcgtttgtt tgttgaagaa gtttgcatta caaaaaaaaa aaaaaaa 887 // ID GT422684; SV 1; linear; mRNA; EST; INV; 840 BP. XX AC GT422684; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-840 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; bc93af18dbd442f732c8da67b8538b30. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 12. XX FH Key Location/Qualifiers FH FT source 1..840 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 840 BP; 314 A; 165 C; 168 G; 193 T; 0 other; aaagtaaagc agatacttga caactaaagg acacccaaga tggtaatgtg ctgggcgttc 60 gagtgtcgtc actacaatgc cagggaaaaa tgcagattct ttagatttcc gaaggatcca 120 gcgctaagaa aaacatggat gtcactagta aggcgtgatg aggaacctgg aacaggttgc 180 tacttgtgtt cgtgtcattt tccgaatgga aaaaaggaaa atttgccact gcttgcactg 240 cacgatgaac gaaaattcta ccccaatttt ccagcagacg atcatagtta ttatgccttg 300 gaaccatcag ccttagagcc agctggacaa aaccccacgc aaaaagacga ttatggcaaa 360 atggtcatgt gccgagtgct caattgtaag cactacaatc aaacggattc ctgctcattt 420 ttcaaatttc ccaaagaccc aaagctaaga agaaaatgga agcaactgtt aaagcataac 480 gttgaacctg gacgaggcga tttcttgtgt tcctgccatt ttcccgatgg aaagaaggaa 540 aaaatgccac tcctgataaa attctgcaaa gaaaaacacc agaggctaaa atcaaagaca 600 cgcacagatg ccaggacctg cacaattacg tctggaggta gtgaataagc caaaataaaa 660 cacatgtaaa agccatgtgc tcaacaaaac attgcaaaag aaaaaacacg ttgacctgaa 720 taaatggaaa atcacactca attttcagtg caatttggag tttgtaaaga tgacatgatc 780 gcaaaagtgt tgtgtttttt aatcaaaata actattttaa caaaaaaaag aaaaaaaaaa 840 // ID GT422685; SV 1; linear; mRNA; EST; INV; 800 BP. XX AC GT422685; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-800 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9bcbc7d4bd86fba2fb8fa48ae60901d3. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 13. XX FH Key Location/Qualifiers FH FT source 1..800 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 800 BP; 266 A; 159 C; 200 G; 175 T; 0 other; gacctgggga atggagttgt tcggttctcc agatccaggg tttaccacaa gaaggctctt 60 tacaagtttg tgggcaagaa agtcgctccc acaaagaaac ccggcaagcc caaagtggtt 120 gtaaagccaa tcggtggaga caagaatgga ggtacacgtg ttgtgctggt tcagaagcgg 180 cgcgaagtgt acccgacgca ggaaaaaatt aagaagaggc caggaaagcg gctgttcaaa 240 aatcatgtaa ggactcttcg tcccagcatc accccaggaa ctgtcctgat cttgctggct 300 ggaggtcaca agggcaaaag ggtagttttc ttgaagcagc ttcaatcagg acttctttta 360 gttactgggc ctttccagat caacgcctgc cctttgagac gcattagtca aagatacgtg 420 attgccactc aaactaaaat cgatatcagt agcgtgaaaa ttgacgataa aatcaacgat 480 gaatacttca aacggcagcg cgaaaagagg gcgaaaaagg aagaaggtga tgtcttccaa 540 aagaagaagg aaggatacaa agtaagtgag gagaggaagg ttgaccagaa ggcagtcgac 600 aaacaggttc tagaaggaat caagaagcat cctgatagga aaattctgtt ggcctaccta 660 tacgctagtt tcgggctcag atccagccaa ttcccacatc gaatgcaatt ttgagcgttg 720 ggcgtaattt ttaaacatgt ttggccctgc tttattttca aatagaacaa gtccatacat 780 agaaaaaaaa aaaaaaaaaa 800 // ID GT422686; SV 1; linear; mRNA; EST; INV; 799 BP. XX AC GT422686; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-799 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 551e095270501d17bd184d48bd2625a6. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 14. XX FH Key Location/Qualifiers FH FT source 1..799 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 799 BP; 270 A; 122 C; 189 G; 218 T; 0 other; gcaggaattg gagaagctat tgcaaagctt ttagtgagaa atggtgttat ggttgctgga 60 gttgcaagaa gagcagaaag cgttcgcgaa catgccaaaa agctgcttgg tgagagagga 120 gttttgcacg catatcaatg tgatctaact aagcaagacg agatattgcg gacattcaag 180 aaaattacca cggaacttgg tccaattagt attttaatca ataatgctgg cgttctcaag 240 gtgtctggta taattgatgg tgacatcgaa aagtggaaag ctgtgataga caccaacgta 300 atggctgtag ccactgggat aagagaagct gttgcaagca tgaaagccca taacatcaaa 360 gggcatatta tcaacattaa tagtactgct gggcatacag ttcttgattt ccctaacgca 420 gggttttatc cagcttctaa atttgctgtt actgctttaa ctgaatcggt taggttggaa 480 atcaacaggg aaaagttgcc aattaaaatt accagcctca gtccggggta tgtggaaaca 540 gatattgtca aagtagcatt tactgaagtg tctggaagta atagttggga gaaaattaca 600 gtgaagggtt tgcacgctcg ggatattgcc gatgcagctt tatatgtttt atcaactcca 660 gagcatgtaa atgttaaaga actgaccgta atgttgcaag gaagtcctgt gtaaaatgga 720 acatgaaata gtatttaaat ggattgctcc tactgaaaaa taataaatat gcttttattg 780 acgaaaaaaa aaaaaaaaa 799 // ID GT422687; SV 1; linear; mRNA; EST; INV; 796 BP. XX AC GT422687; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N15 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-796 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d391a59fc291e27785d394755cbdcf50. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 15. XX FH Key Location/Qualifiers FH FT source 1..796 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 796 BP; 271 A; 114 C; 126 G; 285 T; 0 other; gagcggttat taaaaaggga atcatttgct gtttgaaaat gtttaattgc aatttgtcag 60 aatattattt cagcaaatgc aaaccagtat ttgcactaac tcttacttac gaaaggagca 120 gctgtaaatc tttattgtaa ttgatggtca agagattctt aaagtatctg gtataacttt 180 tctacttctt tcggcttatc gtgtaacaaa gcattgcttt gcataaccag ccccattaga 240 aattgtttta agattgttct tattttgata catagtatcg aaggatttga tcaatttgga 300 atagacaatg cctttggata tgcaacttac ataaattgtg gcatgatggg acgagtattg 360 aatctagtac ttacacagcg aaagcatatt aattttaaat tgaactttca tgcgttataa 420 aatttccact ggaatatcaa tctgacaata tgcaacctga ttgctgactt cctatgcctc 480 attgcactaa tgttcttttt gaatgcactc ttcaaatgga actgaatttt aatttgtttt 540 atgtaattgt gaatcatcta taaactgaac attttttttt atatttcaca gaacaacatc 600 gatatttgac actatatggg taacatttca aaacaattca atttgtacta taatctaggc 660 tttgcgtgta taaattattt tctagataag attttgtttt gtagtataat accgtgataa 720 ctcttaagtt attgaaggta cagattcatt gtgtaaaata gagacaaaaa caaacgggca 780 aaaaaaaaaa aaaaaa 796 // ID GT422688; SV 1; linear; mRNA; EST; INV; 769 BP. XX AC GT422688; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-769 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f73e60e8515a6f305803973db17678b4. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 16. XX FH Key Location/Qualifiers FH FT source 1..769 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 769 BP; 186 A; 245 C; 191 G; 147 T; 0 other; caccttcctg ctgtgcatca tttgcctgtt cgagttcgcg tcggggatcg ccggctacgt 60 gctgaggagc cagacggccg cctatctgga cacgcggctc agggccgacc tgagcaccta 120 caacgagtcg acgcacgcca tctgggacct gatccagagc aatttcgagt gctgcggagt 180 ggacagctac gctgattgga gccagtattt cggcgagagt ggggaactgc ccgtgtcctg 240 ttgtcccagc gtctcgggcg tcgtgggcgc cttctactgc aacagcgcct caacgaccac 300 tggtacccca agtaccagtt actccaccac aacgacgacc acaacccaag gcactacttc 360 aagcgaaacc accacctcaa tcactagctc acccgatccg acacctatat ctgcacggtc 420 tgatcaaaac cagttagtga acctggcggg ctcagagtcg acgacagccc cctacacaga 480 gggctgcaaa agcgccttcg gcgcctacat caagcagcac gcctaccaga tcgggggctg 540 cgccctgggc ctggccatcc ttcagctggc cggcatcggc ttctccttct acctggtgcg 600 ccagctgaag aacggctact tcagcacctg atcaccggcc gcggcccctc gcccatgtaa 660 ataaccccta gttagtggcg tgtttttttg gaaaatactc ttataatgaa gcggaaatca 720 accaaccaat taaacacgta gtgagacact gtaaaaaaaa aaaaaaaaa 769 // ID GT422689; SV 1; linear; mRNA; EST; INV; 857 BP. XX AC GT422689; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N17 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N17 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-857 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 81f981cb67c5359c38d2a2b4d87c9ec8. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 17. XX FH Key Location/Qualifiers FH FT source 1..857 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N17" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 857 BP; 239 A; 187 C; 175 G; 256 T; 0 other; caacaagcat ccctttaaat tgattaaatt ccagaatggg agattttggg ggtcaagagt 60 tctactggaa caaccaggct gaagtacctt tacaacaagg ctattacgaa catgattttc 120 agcaattttc caaccagcaa ctagaattca acccccaaga agatttcgtc aatcagcagt 180 ttgctcagta tcaaccaacc actttttaca accctaatga ctatgacacc agcagagaca 240 ctgacgagga gttcgtagag cccccgctgt tggaggaact ggaaatattc ccagaccgga 300 ttctggagaa ggttttggcc gttctgaacc cactcagagg acatagttta gcagacgatg 360 ccgaatattt gactaaagac gccgacttgg ctggtccggt gttcttctgt ctggccctgg 420 ctatgctatt gctcttagca ggaaagtccc aatcctttgg atacatttat ggattttcca 480 tgatatcgtg catcttaaca tactgtcttc tttccctgat gtccagtgct gaaaagacac 540 ttacctttag cacagtcgct tcaattctgg gctattgctt gatcccaata gtggctttat 600 cgttccttgg cgtgtttttc aagctaagcg gactcgtggg catcatttta gctggctgtg 660 ctgtcttctg ggccagcttt tcagcctcca ggttgtttgt tgcagtttca ggcgataagc 720 aacagcagcc tttaatcatg tatccctctg ccttggtgta tggagttttc gtattattgg 780 tgttgtttta aatagaaatg tatatacaat taaatattta acaccgttaa aaaaaaaata 840 aaaaaaaaaa aaaaaaa 857 // ID GT422690; SV 1; linear; mRNA; EST; INV; 757 BP. XX AC GT422690; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-757 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4d7c092f8c2c8b85f0ba6ba616bc3852. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 18. XX FH Key Location/Qualifiers FH FT source 1..757 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 757 BP; 235 A; 167 C; 197 G; 158 T; 0 other; ggaggatcaa ggcctgggcc gatattcaaa aggtctacaa gagtcaacgt ttccgtgctg 60 gcaagggaaa aatgcgcaac aggaggagaa ttcagcgtaa gggacctctg gtcgtttatc 120 atcaagatca aggtctacgc agagctttcc gcaatatccc gggtattgat ttaatcagta 180 tcgagaagtt gaatctactg aaattggcgc ccggcggtca tgttgggcgt ttcgtcatct 240 ggacggaatc cgctttccaa aagctggaca aagtgttcgg aagttggaaa gtggcttcca 300 gccacaaaaa gggctacaat ctgcccgaaa ctaaaatgtc caacaccgat ctgtctcgcc 360 tgttgaaggc tgatgaaatc aaggcggtcc tgcgtcgtcc acagaagaag attgtgcgtc 420 gtgtgcgacg cctcaatcca ttgacgaacg ccaaggctat gttgcggctc aacccgtact 480 ctgcggtttt gaagcgcgaa gcaatcctat caggccagaa aagaagcttg gctagagagg 540 aacttcttgc caagaagcgc gggatcactt taccggaaac aagtccggta atcaggtcgg 600 ccaagctgca ggctaaaagg agggtccaaa tcttgaaggc ccagaaggca aaggctgcca 660 agccgaaaaa agttgccgcg aaggcaccag ctaagaaata aatataatgt ttcttagtta 720 cgaaataatt atcaagtaag aaaaaaaaaa aaaaaaa 757 // ID GT422691; SV 1; linear; mRNA; EST; INV; 837 BP. XX AC GT422691; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-837 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3a20adddf1caae354687c5b33680f357. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 19. XX FH Key Location/Qualifiers FH FT source 1..837 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 837 BP; 269 A; 163 C; 190 G; 215 T; 0 other; ctgtagagcg cctggctcag agagttgcaa ctgaaaaagt ggtcgtttcg tgtgtgccca 60 catccttcca ggcaaggcaa ttgttggtcc aaaaccagtt aacccttacg gatctggacg 120 tgacccccaa cttggacgtt tgcattgatg gtgctgatga agtcgattct aaccttgatc 180 ttatcaaagg gggaggtgga tgcctgctgc aggagaaaat tgtggccagc tgcgctcgca 240 agctgataat catagcagac tacacaaaaa acagctctca actaggtgaa cagtacaaga 300 aaggcattcc aatcgaagtg gcccccatgg cttacttccc aataaaaact cgccttgaag 360 caaagtatca agggcaattt atgctgcgaa tggcggtgaa taaggcaggc cctgttgtaa 420 ctgataatgg caattttatc ctcgattgga caaatttcga ccaaacctta gaatggaaaa 480 acgttaacca ggaactggca ctgatccctg gagtggtaga gaccggatta ttcatccaaa 540 tggctcagga ggcctatttc gggatgccca atgggagcgt gaaggttcag aagagatctg 600 ctgctaatag cttgccattt ttttcatctg actaaaaaat atatagacaa aaccattgtg 660 atatattgaa cacatatttg tagacaccca gatacttcaa ctttagttaa gtttaaaata 720 ttacaaaggg gcttttatag tgattgtacg gggcgacaaa gttatttttg taaaaaatgt 780 attaagtgta tcaaaaatcg gtgaatatat cgtggtaaac caaaaaaaaa aaaaaaa 837 // ID GT422692; SV 1; linear; mRNA; EST; INV; 820 BP. XX AC GT422692; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-820 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2e4ffdfa0a5d72d1832c18f7216db865. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 21. XX FH Key Location/Qualifiers FH FT source 1..820 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 820 BP; 269 A; 168 C; 195 G; 188 T; 0 other; cagcgacgcg gacgttcaaa aacagatcaa gcacatgatg gctttcatcg aacaagaggc 60 caacgagaag gctgaagaaa tcgatgccaa ggctgaggaa gagttcaata ttgagaaggg 120 ccgtttggtc cagcagcaaa ggctgaagat catggaatac tacgagaaga aggagaaaca 180 agtcgaatta cagaagaaaa ttcaatcttc aaacatgttg aaccaggccc gtttgaaggt 240 tctcaaagta cgggaagatc atgtgcggtc tgtactggag gatgctcgta agaggctggg 300 tgaagtcacc agagatcagg gacgttacgc ccaaattgct gagtctttga ttttgcaagc 360 actctaccaa ctgtttgaaa acaacgtaat tgttcgaacg agaccacaag acagggactt 420 ggtgaaaacg gttttgccaa cagtggctac taaatatcgg gatgttactg ggcgcgacgt 480 caacgtaacc cttgatgatg ccgtccagct ttcccaagac atcactggag gaatcgacct 540 gtacactcgt caaaacaaaa tcaagatcag caacactttg gaggcccgtc tggagctcat 600 ttcccaacag ctcattccac agattcgtaa cgctttgttc ggacgcaacg tcaaccgcaa 660 attcaccgat taaattgctt tattttggaa gcaaattatt gtaggaagct cttggaacag 720 atgtgcaatg gttacaggtc tttgctggct agtttcaagg cttcaagtat ttctagtgtg 780 aaaaaaatat acgctaaatc agtaaaaaaa aaaaaaaaaa 820 // ID GT422693; SV 1; linear; mRNA; EST; INV; 842 BP. XX AC GT422693; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-842 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4fead5b31360799de5914d5369eaa550. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 22. XX FH Key Location/Qualifiers FH FT source 1..842 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 842 BP; 237 A; 167 C; 186 G; 252 T; 0 other; cgaattgtta caagaaaccc ccaaagaagt ggtcatggtt gagctggacg aggtcgtgat 60 taaggcctgc tccaaataca tgagatccgt ttgcggagac gcgatggaca actacgatgg 120 acctcatcat aagataataa ttggcgattg tttgccggtt ctcgaacagt acatcggcga 180 gggcagacgg ttcgattatg tttttggcga tcttactgac gtgcccattt cggtcgacaa 240 atctttggaa ttgtgggcct tcattaacaa ggtgctgaaa ctgagttttc aagtcttgaa 300 gcctgatggg aaattcatga ctcacgtgac tggccaaggt tgtaccgaag ctttggaggc 360 ttataaacaa tgcctggatc gactgcagcc tgcagtgaaa tacgaaacgt cctcaaattt 420 cgtgcccagc ttccacgaga cctgggtttt ctgtcaagtt agttttaaga acggcgaagc 480 agcgacaagt cagcaggccg aataacgata cgtttcgatt tgcctatttg ttacttcaat 540 tagtctagat ctgttactag atcagaacgg ctcacatgag tctggactaa ctattcttta 600 tcaattaacg agaactagtt tagtccccca gccccaacca agttgctttt tgccataagc 660 tagagtcagt atttttctac aaatgtgttc ctcgcttcgt tccgaaaatt tgcgctatat 720 ttagatctta aggagcaatt cgtatgtaag caggtttgta ttttcataat ggttattatt 780 atctgtttaa tttttgtttt aaatgcacgg tacctttatg aagtaaaaaa aaaaaaaaaa 840 aa 842 // ID GT422694; SV 1; linear; mRNA; EST; INV; 760 BP. XX AC GT422694; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-760 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; b560b4f53c280194ea5d7aaaa5df4d7a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 23. XX FH Key Location/Qualifiers FH FT source 1..760 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 760 BP; 221 A; 151 C; 178 G; 209 T; 1 other; gggggtccag tcatcctgac aatctagaac tgacgccatt tntcctgcat tcgacttaaa 60 aagtgcaagt gatttaatca aacctctgca aagtttccaa aatggttaaa gccgttgccg 120 tgttgaaaag cgaagtggtc aatggaaccg tcttcttttc ccaggaaggc aacaatccgg 180 tgcaggtaaa tggatcgctg agcggtctca aagaaggcct gcatggattc cacattcatg 240 aattcgggga caacaccaat ggctgcatat cagccgggcc ccatttcaac cccaacgaca 300 aggaacatgg aggcccaact gacgctgacc gtcatgctgg agatttagga aacattgagg 360 ccaatgctga aggagtcgct aagatcaaca tcacggacaa gcagatttcg ttgagtggag 420 cgaacagcat tattggacgc acagttgttg tgcatgcaga ccctgatgat ttgggcaagg 480 ggggacatga gctgagcaaa accactggca atgctggtgg caggttggcc tgtgcagtta 540 ttggccttgc acagcccaac tagatttatc aagtcactgg tagtttacgt taagatgcac 600 tgttcttttt ggtggtattg agaatttctt tgtttttagg aatcgttcct atttcctgca 660 ttttaggtgt tatatattga aaatgattca ttttatatta tgttcttgtt gttcatttca 720 gtttaaataa aaaaatacta tctaaaaaaa aaaaaaaaaa 760 // ID GT422695; SV 1; linear; mRNA; EST; INV; 792 BP. XX AC GT422695; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_N24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_N24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-792 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; b132a2b062854f0061d89cfa90a6b363. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: N column: 24. XX FH Key Location/Qualifiers FH FT source 1..792 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_N24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 792 BP; 264 A; 146 C; 171 G; 211 T; 0 other; aataaaaaaa ataataaatc ctgctttttt gcctccgatt gtatgaaact gactgactga 60 aacgagttgt ttcagctgat ctttctatgg agaactttgt ccaacatggg accaaatgca 120 ttggcgtggc cgctaattat aggtctcttc taaaagtcct taacagatca acgccagatg 180 ttccagactt tttcatgaag ccccctagtt cttatataac ggaaggacaa aaaataatta 240 tacctaaagg tttctctgtt aacgaagaaa tcgaactggg agtggtaata ggaaaacatg 300 ggagaaaagt gtctgaatgc aaggcaatgg atatagttgg tggatactgt ctagctttgg 360 acatgaccgc tacttgcaga cttggcctag ccagacaaac tggtggaagt tggacattag 420 gaaaaggttt tgatacagcc tgtcctgtga gcaaattcat cccaaaatca caaatacccg 480 atccgcacaa tgtggaatta tggtgcactg taaatggagt ggaaaaacag cgcggcaaca 540 ccgacgactt agttttcaat attcccttcc tgatttcgta catttccgaa tatattacgc 600 ttgaaccgaa cgatgtgatc ataacaggat ctccgcctgg aatggggcct gttacgaagg 660 gcgacgttat agcaggaggc attactgggg gtgagagcat gaaatttgat gttgatgctg 720 aataaacata gtaatacatt attatcatca ttattaataa agaaagccgt tcacaaaaaa 780 aaaaaaaaaa aa 792 // ID GT422696; SV 1; linear; mRNA; EST; INV; 318 BP. XX AC GT422696; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-318 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2731314358431fd4f153daf25b42e0ba. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 1. XX FH Key Location/Qualifiers FH FT source 1..318 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 318 BP; 107 A; 49 C; 67 G; 95 T; 0 other; gagatgttat gggctcgttg gacgttgtta tgcttactat agacgaaatt tgcgatagtg 60 gaatcatcag tgatgcggaa ttgtagctcc gtagtctcaa gagtcgcagt gagaagtgat 120 gacatcccaa ttggagagca aactgttgct caggtgtttc aaactgcaaa agagcagctc 180 aaatggtcat ttcttaagtg agttcaaaga gctcagaatc ctttttctgt ataatcttgg 240 tatttatcat caagccagaa tgttccgttt aatgatacaa tttaatatac tttacaaata 300 gaaaaaaaaa aaaaaaaa 318 // ID GT422697; SV 1; linear; mRNA; EST; INV; 412 BP. XX AC GT422697; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O02 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-412 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 7322b4045c6b46b0e708d958efe4ce4d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 2. XX FH Key Location/Qualifiers FH FT source 1..412 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 412 BP; 137 A; 102 C; 75 G; 98 T; 0 other; gggggaaccc ggcaacattg gtcaactcgt tgtgtaactt aaagtcgaag agcaaatttt 60 ccagttttcc ttgcagcaaa agagcggaaa cacaaaccac catgtcagac acaccagccc 120 ccgctcgcca catgaagtac ccatacactt tgtcggctaa actagcccaa tttccataca 180 agttctacct aaaaaataac tgggttgtta agtactacct gatttcgctt gttgtatgca 240 ttcccgtctt caagtcgatc agcaacttgt cgaattcacc tgcaaatgtt gcaaaatggg 300 ccgaaattcg ccgcaaggaa gccgagggac accattgagc gctctaatgc ctttaatccc 360 gatagaactt ttaaatatac ctaaattata gagccaaaaa aaaaaaaaaa aa 412 // ID GT422698; SV 1; linear; mRNA; EST; INV; 856 BP. XX AC GT422698; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-856 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a900cbb0cb1590b94739fc9b5380d00b. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 3. XX FH Key Location/Qualifiers FH FT source 1..856 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 856 BP; 289 A; 132 C; 182 G; 253 T; 0 other; aatgatggag ttaaggcggt gcttttaggg ccccctggtt caggcaaagg aacgatggcg 60 gcttggttga aagataaatt tttcgtttgc catctttcaa caggagatat gttacgagct 120 gaaatatctt ccggatccga attgggatcc aggcttaaaa aggtcatgga tgaaggaaag 180 ttggttagcg acgatttggt cgttaatatg atcgacaaga atctggataa gccagaatgt 240 aaaaatggat tcctccttga cggatttcct agaactgtgg tgcaagctga aaagcttgat 300 aatttgttgg ataaaagaaa tacgcaactt gatgcagttg ttgaatttgg aattgaggac 360 agtctattgt ttcgccgaat cactgggcgt ttgattcatc cagcaagtgg cagatcatat 420 catgaggagt ttgcaccacc aaaaaaaccc ttgacagatg atataactgg agaacctttg 480 atcaaacgct ctgatgataa tgtagaggct ttaaagaaga gattggcaac atatcataac 540 cagaccaaac ctttggtgga ttattatcaa attagaggta ttcattaccg cgtcgacgct 600 gctcaagcag ccaaagatgt gtttaagaac atagacaata tttttataaa aaaatctgca 660 aagaacgtag cctcgagact ttaaaagaag tttattttta gattttccca tccttaaaaa 720 cgtatttgca tttttagtta cttttttgtt taaaattcaa acaaataaaa tcggtgttaa 780 ggcataccaa attggaaact ttttgtgtga ttctggttta attaaaaatt gcattaatga 840 aaaaaaaaaa aaaaaa 856 // ID GT422699; SV 1; linear; mRNA; EST; INV; 879 BP. XX AC GT422699; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-879 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 768b279e3d6541fcf318a62d4ccf3e48. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 5. XX FH Key Location/Qualifiers FH FT source 1..879 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 879 BP; 259 A; 139 C; 150 G; 331 T; 0 other; accgcacatg attgttgtaa aaactataat tgcttgtttt tagtcaatat tagaacattc 60 ttagcccttt aggcccccag acgtttccac gtagcttatt tgaaatgtat aatagggaat 120 atattttagt atatttttag aaccagcgtg cacatgcgaa tagaagaatg tatttgtcga 180 ctgagaatat tcatgttcaa gttcttctat cagtattttg atgtgtgcgc caactaggag 240 atctctatag taattttagc gattagtttt acattgtata atccagtttt agtgtaggaa 300 tcaatatgac aactttccta ggaaatttta cgaatgctct tgtctttttc ttccactcgg 360 ttcttaggcg gttggtttat cttggattac atccacattc catttgtatt ctgtagacgt 420 tgaaaactta ttccattatg ttcagtattt agactatgcc tatgtttgta aatattcttt 480 aaggtagatt tacttgtata atactttcgg attagctgtt gatatatctt cataagtttc 540 ccgtgacgga ccactgtttt attttttata ggccctaatt tatacgtgta cttaggagaa 600 tcaaattgtt attatctaat gatttttaca gacatacgag agtttatatt gcgagtatat 660 ttaccaaatg ttttgcaaag ttgtttgcca atccggtata attcatcggg aaattgctcc 720 agtcgatttc aaccttaata cagttttttg tgtataaata tgcaaccaaa accagcagtt 780 tgggcgctcg gcttggagag tttcacggtg aactctgtca tattgcaaat gattttccag 840 acaatatact taatagttta taaaaaaaaa aaaaaaaaa 879 // ID GT422700; SV 1; linear; mRNA; EST; INV; 815 BP. XX AC GT422700; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O06 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-815 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 51280171aa476afbb5b67c0326e460d5. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 6. XX FH Key Location/Qualifiers FH FT source 1..815 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 815 BP; 273 A; 159 C; 149 G; 234 T; 0 other; gaaatatatt caaagttccc tgaagcaaga agatgatata caattaccaa caccatggaa 60 gcaaatcttc acctgtgtgc cactatacgc tatactcatt gcatatgttg gaaatctatt 120 tggatatgcc gttatgacca cggaaatacc aacttattta gccaaagtta tgagtttcga 180 tgttaaaagg aatgcaataa ataatgcaat tccaaccgct gtctctctgc tagcttcaat 240 gatcaccggc cctgcatcag actggcttat ccaaaaacaa tatttatcta ctttaaacac 300 cagaagattt gcgcaaataa ttggatccta tgggaatgca atctgcctta tctggatgtc 360 attcctgtct aaagaccagg cctatatggc agttataata ttgtcttttg ccagtacttt 420 aacatcgttt gttcaaattg ggtgcaatgt aaaccacctg gatttgtcac cgagatttag 480 tggagttatt tttggtatcc tcaacgccat tggtcaatta gcttcagtag tagctccatt 540 atttgtacag tttgctgtta cggatacgaa aagcatcgag caatggcgca cagtttttat 600 catcacagct ggatttttta tcacaggagc atcggtattt tacttcctag catcagccac 660 aagacaaacg tgggatggac ctgatctgaa agatctgtcg gagaataaaa aaaaaacagt 720 ccagtgccga acgtgagaca agaagcagtt aaattggcaa ctgtacttaa ataagtaata 780 aactgaaatt aacacagtaa aaaaaaaaaa aaaaa 815 // ID GT422701; SV 1; linear; mRNA; EST; INV; 817 BP. XX AC GT422701; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-817 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f3ef4631e2f0db4c95b9a7ae12ab46be. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 7. XX FH Key Location/Qualifiers FH FT source 1..817 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 817 BP; 278 A; 155 C; 199 G; 185 T; 0 other; atgcgatcga cctgcaggta ttgccagggc acgagaatgt acattaagga taaatgtatc 60 gaatgtgaag gcaaaggctc tacagttcag agacgcacca tcgcgattcc agttccagca 120 ggtattgagg atggccaaac tgtacgcatg tcggtgggac tcaaggaact gttcgtcaca 180 tttcgcgtgg ataagtccga ttacttcaaa cgtgatggtg ctgacgtcca tacagaggcg 240 gctatttcag tagctcagtc cttactagga ggcagcataa gaatacaagg actctatgaa 300 gatcatgtgc ttcaaatcag gcctggtacg tcttcacaca cgagaattag gctgtcaggc 360 aagggtttga aaaaagtgaa tggctatggt aacggcgatc attatgtgac tttgaagatc 420 aaagtgccga gccacttgga cgagaagcaa aaggctctag cccaagccta cgctgaactg 480 gaacaagaca ctcctggaca gattttcgga attacgtaca acaccgatgg tgaaaaagcg 540 atcagttcga gcaaaaatca ggctctgttg gcggcaatca gatcagccct ggaagggaga 600 aacaagtaaa aaataaaccg gacaacgtga aaagtgccac cgacgaagag cacagcgatg 660 tagagcatga aaaagtgcaa aagaaaattt aagttttgaa aaaaattgcg atcggttcag 720 ttattccaaa gaataaacgg gattgtgtat acttgcagtt gttacatttt attaaattca 780 tatattgctt tgaatagatc aaaaaaaaaa aaaaaaa 817 // ID GT422702; SV 1; linear; mRNA; EST; INV; 810 BP. XX AC GT422702; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O08 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-810 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6b481b04e017ef8642b98fa097a122ee. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 8. XX FH Key Location/Qualifiers FH FT source 1..810 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 810 BP; 272 A; 164 C; 199 G; 175 T; 0 other; gagcgaaagc gccagatcga ggagaatacg atacgcagca cccaacacag tctgagcatc 60 ctgcgggact ctgaacagat cggaattgcc actgctgagg agctgatgag gcagcgggag 120 aagctggaga agaccgataa gcagctggac gacatcaaca acacgctcag gttttcgcag 180 aagcacatca atggcatcaa gtcggtgttc agtagtttga agaattacgt gtctgggagg 240 aacgataaga gccccactac gtccagctcg acgcctagca ctgttcggga atccacgtcg 300 aattcaaatc tgcaccagaa gctgcagact tatgatcgct atgatgagca tcccgttact 360 cggcttaggg gcaacgacta cagtatgcat cagcaacaaa cgacgggatc aggaagcagg 420 gatttcagtg caaaattaga cgcaaacttg caggaaatgt ccagcaatat atgcaggtta 480 aaaggactgg catgcgaact gggacaggaa atcgactccc agaatgatct cattgataat 540 atcacttaca agaccgacca ggccgatttg accattcaaa ggcaaaacaa agacatggtg 600 aagattttga aaaaataaac cagttcgatt atatatgagc gtaaatatgg catgtggcca 660 aaggtcgggg gctgcaaaca gaatttgatt ggtggaacag tacaaaactg ccggagtttt 720 aaagtctatt agtatttatg aatctatttg tatattgtgc atgatcgaat aaactaattt 780 gcattgtttt ctgaaaaaaa aaaaaaaaaa 810 // ID GT422703; SV 1; linear; mRNA; EST; INV; 832 BP. XX AC GT422703; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O09 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-832 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a919bbd53e22b0bd773eef6b07ca0095. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 9. XX FH Key Location/Qualifiers FH FT source 1..832 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 832 BP; 225 A; 169 C; 185 G; 253 T; 0 other; cagccgctct accggccaac actattactt gaacatctac acaaaggagt cgcaatggga 60 cgctccaagt gagcctgctg aacgcggtca gtcttctggg cctgaccaag tccagtgttc 120 ccatctgtta gtcaagcaca aggactctag gcgtccgtcc tcatggaggg aagaagtcat 180 tacacgctcc aaggaggaag ctttggagct ggtgaaaatg taccgggagc agatcgcgca 240 gggaaaggca tctttcgctg aactggccac aaaatattcg gactgttcat cggctaagcg 300 cggcggagat ctggggccgt tcggcaaagg agccatgcag aagccctttg aggattcggc 360 ctttagtctt aaggtgggcg agttgtcaga tccagttttt acagattcag gcgttcatat 420 tattttgcgt accgtttaaa gttgtagcgt tcaaagcgaa aagtattcat ttttttgtct 480 aattgttaat gaatttaaac ggcggttagg cgatagttta tcatagtaat tcctggtagt 540 aactagctgt ttcttcatgc aaatttgaac gttttccacc gttcctcagc tcagccaaag 600 ttactgtggt gcttgatatt tctctttttt ctcaattatt tcagttagtt attaagcaat 660 tatcacatgc ggtaatttcg agcaccttgc tagctcttta ttaaggcccg aattttacca 720 gcactggaga tacaatttat tatgacagat tttcaattga ttgtttttta attattgcaa 780 ttaaataaac atttttcagt tccttgttgg caatgaaaaa aaaaaaaaaa aa 832 // ID GT422704; SV 1; linear; mRNA; EST; INV; 795 BP. XX AC GT422704; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O10 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-795 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 389659cbe11cc52732952afbbf79b6e4. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 10. XX FH Key Location/Qualifiers FH FT source 1..795 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 795 BP; 244 A; 182 C; 196 G; 173 T; 0 other; cccatcgagg gagcggtgga aattgacggt ctcaacaccg gaaaagtggg cctgagggcg 60 ctgcgctctt caatctccat tatcccccag gtgcccacac tcttctccgc cagtctcaga 120 tataacttgg atccgttcga aaaatgcgca gacgatgagc tctggaaggc ggtagaacgt 180 gtagagctga aacagagcgg cgttgcgctc gacactgctg taagtgaaag tggggccaat 240 ttcagcgctg gccaaagaca gttgatttgt ctggcacgag cgatagttag gaataacaaa 300 attctggtaa tggacgaggc caccgccaat gtggatcaac acaccgactc cctgattcaa 360 accaccatcc ggcaggcctt caaggattgc acagttataa cgatagcgca tcgcattaac 420 acgataatcg actgcgatcg ggttctggtg atggatcatg gccaagcggt ggaatttgcg 480 gcgccccacg agctgctgga gcagaaagat gggcattttg cccaaatggt tgaacaaacc 540 gggccgacca tgtcgtccaa gctacgcaaa caggccaaag atagctacaa tcagaagagg 600 gccgtggaga atcaaaagag cgaataactg acttaatgga aaataattaa cgctgcatgg 660 ggaacattcc cagacaataa caccgactga ctatttactt gatcttaaat gcttacttac 720 tgttttacgg atacttacaa ggaatactta atatttgtga ataaaaagtt tgtttttcaa 780 aaaaaaaaaa aaaaa 795 // ID GT422705; SV 1; linear; mRNA; EST; INV; 767 BP. XX AC GT422705; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-767 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9019a8f4d585cc517218e1c664354808. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 11. XX FH Key Location/Qualifiers FH FT source 1..767 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 767 BP; 265 A; 129 C; 127 G; 246 T; 0 other; cctttagatt ttacttttgt ttgcccaact gaaattctcg cgtttagcga tagaatagga 60 gagtttaaaa aaatcgacac tgaagttgtg gcctgctcag tggattccca atttactcat 120 ttagcctgga ttaacacgcc cagacaacag ggtggtttgg gcaaaataaa tatcccatta 180 ttatcagatt taactcacag tatatctaaa gactatggag tatttttaga gaatgccggc 240 catactctaa gaggtctttt cattatcgac ccattgggaa ttgttaggca aataacaatg 300 aatgatttgc cagtcggtag aagtgttgat gaaaccttaa ggctagttca agcttttcag 360 tataccgaca aacatggaga agtgtgccct gctggatggg taccaggcca agatactata 420 attcctgatc ctatcgagaa gcagaaatac ttcaaaaacc actagtgttt ttgcatttac 480 atatcgggca ttttgaaaac ttgggaacag ctcaaactca tgacaattta ttacatttca 540 gtggtaggca agttttaaac tatattaaca gtttttatct taagaagata tatatctttg 600 taataattga caactagtaa ctttctttat taagatctca ttctctgttt tcctcccaag 660 acatcacatt ttatttttaa tttagtatta aaggcaaatt atttttccat atgcaaataa 720 aactgtgctt ttaaaaaaaa aaaaaaaata aaaaaaaaaa aaaaaaa 767 // ID GT422706; SV 1; linear; mRNA; EST; INV; 798 BP. XX AC GT422706; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-798 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3ebe79d5895c76494ffe8bfe04803962. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 12. XX FH Key Location/Qualifiers FH FT source 1..798 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 798 BP; 255 A; 141 C; 192 G; 210 T; 0 other; ctacggattg tggaggagct gaaggaaaat gcctgtacat agacaccgaa gggacgttta 60 ggccggaaag gttacttgct gtagctgagc gctttaaaat ggagccccag actgttttag 120 ataatgtcgc gtatgcacga gcttacaata ctgaccatca aacccaactg ctcctgcatg 180 ccagcgccat gatggcagaa tcaaggtatg ctctattagt ggtggacagt gcaatggctc 240 tgtacagatc cgagtactcc ggaaggggtg aactagctgc cagacaaatg catttatcca 300 gatttctgag aatgttgctc agattggctg atgagtttgg tgttgctgtt gttataacaa 360 accaagtcgt ggcccaggtg gatgggactg caatgtttaa cgctgacccc aaaaagccta 420 ttggaggcca cataatggct cactcatcca caactcgatt gtatttaaga aaaggtcgca 480 atgaaaccag aatgtgcaaa atttacgact cgccttgtct gcctgaagct gaagcgatgt 540 ttgcgataaa tcccgatggc gttggcgatg ctaaggaata agaaaatgtg tggactaagt 600 agttaaggcg gaattatcgt tatttttgag ataggaattg tttcaagttc cataagaagt 660 ggctttatga agcaaattaa tttaaattta ggttaactgg agtaaatatt taatagaaat 720 agaaaacgac actaatttga gctggtgcac tgtaatagga caaataaatc aattgtttta 780 caaaaaaaaa aaaaaaaa 798 // ID GT422707; SV 1; linear; mRNA; EST; INV; 855 BP. XX AC GT422707; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-855 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 468c1454abff118a4b925c5b4d5675e2. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 13. XX FH Key Location/Qualifiers FH FT source 1..855 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 855 BP; 269 A; 155 C; 172 G; 259 T; 0 other; atacggttgg ggtgttttgg cccagtttgg aggagagcaa tttgctgctg aagtcgctaa 60 aatagtaacg actctcgtgg aagtgattaa tgatgcagaa tcacgagatc tgcgtaatat 120 caacgccaca gaaaacgcaa tttcggcggt ggcgaaaatt ttaaagtaca acaacactca 180 aatcaatgtc aatgatgtta tcccgctctg gttctcgtgg ttgcctgttg tcgaagacgc 240 tgatgaagca gtacacgtct atggatatat gtgcgatctc attgagcaga ataatctggc 300 tatcataggt gcgaataacg agaatattcc caaaatcgtt cacattatcg cagagacgtt 360 tcaccgagag gcggtagaaa ccaacacacc agtagcagtg agaatgacaa acatagtccg 420 acaggtgcaa tcaaacgaaa cgttttttca atcactgatt cacacaatga ccactgaaca 480 gcaacaggct ctgcatttcg ctttgcatgc aactcctgcg tcttagttaa taagctattc 540 ctatttcatt gttttacaat aacgtcaaaa attacggttt tattaatata caatatatgt 600 tattttttta aacagtaagg tgttattcgc agggacgcgc gacgcctttt agaactgtta 660 attgtctgta catagtttgc tcgcaaactt ttctgcatgt gatatttaat tttcaatcct 720 atgggcaatt gtctgtgttc ggaaagttgt tttctattaa tgttgtaaac ggagaaatcg 780 tcttgattaa ataaattaat tttctctaag ggggttttta gtaaaactat tatctagcaa 840 aaaaaaaaaa aaaaa 855 // ID GT422708; SV 1; linear; mRNA; EST; INV; 704 BP. XX AC GT422708; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-704 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4d45a5ebeaaf05d8d9ae41a98637b9fe. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 14. XX FH Key Location/Qualifiers FH FT source 1..704 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 704 BP; 200 A; 155 C; 201 G; 148 T; 0 other; gggggctcag tgtgggaata tcagcccgat tctaacaaat aagaggcgga ttttgtgtga 60 gtgatttcaa tcaagatggc tttggaagcg gttttaggca aaaaatacaa gctggccagc 120 agcgaaaagt tcgacgaata catgaaggca ctgggggtgg gcctggtgac gcgcaaggtg 180 gggaatgcgg tgtcgccagt ggtggaccta cgcaaagagg gggacgagta cattcttacc 240 tcgagctcca ccttcaagaa tatggtgacg aaattcaagc ctggggtgga attcgacgac 300 gaaacggccg acggtcgcaa ggtcaagacc actttcacgg tggaggggaa cacgttgaag 360 gaggtgcaga aatgccccga tggcaaagtg accaccatcg accgcacctg gactgatgat 420 gaagtcaaaa cgattttgat tgtgggcgac atcacatgca ccagaatcta caagctgcaa 480 gcctaaaggc gattgttctc gagtgcgaca aggacaccac atcgaactga ttttgctact 540 actacggaat tcacagtttt ttaagatttt tactcaatgt agtcgggggc gtggctctct 600 ataggctccg tccacgttaa cggggcggag cctgttcagg cccacgcccc cttgcgggtt 660 ggatgcccaa aaacaagctg tttcactaaa aaaaaaaaaa aaaa 704 // ID GT422709; SV 1; linear; mRNA; EST; INV; 796 BP. XX AC GT422709; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-796 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f450f52739ae82d80d9a5bcb8ce30561. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 16. XX FH Key Location/Qualifiers FH FT source 1..796 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 796 BP; 279 A; 117 C; 174 G; 226 T; 0 other; ggttggagta gtattgcgat aataacgttt gtgaatatac cttgtcgatg ttcgaagaaa 60 ggaggccagt actggaatta ccaatctgct tttaaatcgg gcttgagtgt ggaaacaacg 120 acacccatct tcgactgtat ccagaagtag ctggacgcat gaaagcaggc cgcaattgaa 180 actagcgaac agttttcagt gaaaatccaa tttaatagta ttgtttttat atttacggat 240 agcgataaat tgtgagtcaa aagcatgaac aagatagaat tagaaggaac agcccatttg 300 cttgatgagc ttgacaaaaa acttatggtt ttactgcgcg atggtaggac cttaataggc 360 tacttgagaa gtgtagatca gtttgcgaat ttagttctac ataaaactat agaaaggatt 420 catgttggca aggaatacgg tgacataccc cggggaattt ttattgtacg aggagaaaat 480 gttgtgcttc taggcgaaat tgatatagac agagaggata aattaccgct aacagagatc 540 tcggtggatg atgttttaga tgcacaaaga aaagaacaag agttacatat cgaaaaacag 600 cggcttattt caaaagctct aaaggaaagg ggtctgcaaa tgaatgcgga tttagttcaa 660 gatgatttat attaaaattg gaactgacga gcacactatt ttattttctt tttataacta 720 ggccccatta aatactggta taaatcgccg ttctgttgaa taacatgcta tttatttcaa 780 aaaaaaaaaa aaaaaa 796 // ID GT422710; SV 1; linear; mRNA; EST; INV; 803 BP. XX AC GT422710; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O17 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O17 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-803 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c69ae9c407aff684508c653cfa9aa47f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 17. XX FH Key Location/Qualifiers FH FT source 1..803 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O17" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 803 BP; 243 A; 129 C; 151 G; 280 T; 0 other; actggactct ctattaaccg atgaaagttt agcaaattgc ccagtgttga ttttaggcaa 60 caagatcgat ttgccctctg ccgcttcaga agatgaactt agaaattatt ttgccctgtt 120 tggacagaca acgggaaagg gaaaagtacc aagaacagac cttcctggcc gtccactgga 180 attattcatg tgctcgattt tgaaaaggca aggttacggc gaagggttcc gctggttggc 240 acagtacatc gactgagtca ctaatataat ttatttgtcc cgtgaaattt tttttttgtt 300 aagaaggaac gtgtcaaaat ctgatcgaac accacacaaa aaaaattcgg aacgttatag 360 gtaattattt atttaaagcg tttctttccc ataaattgtt tccgtttatt gttaaagctt 420 ccccgttgac tagtcttgtt gaaatagtgt taggtagcta aacgatttta tctgtttgtg 480 cggggtgttt atataactgg gtttatacga tcatttcagc tcgcttagcc atatatttga 540 cttttttgtt atctataatg tttgttaatc ctatgaaggt taaaaaagta atagaagcac 600 acgttctggt cttcaacagc gctgcttaaa attttaaatt gctattacat ttgtatgtaa 660 cgagtttttg caattattac ttgagtaatt gctgttacct tgttgataat tgggggaagt 720 cgacaactga tattacacat tcttttaaat atacttgtac ttttttgata ataaattgtg 780 ttctcaaaaa aaaaaaaaaa aaa 803 // ID GT422711; SV 1; linear; mRNA; EST; INV; 595 BP. XX AC GT422711; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-595 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; edf2d24170dbd44a447fe96a23433a81. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 18. XX FH Key Location/Qualifiers FH FT source 1..595 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 595 BP; 180 A; 134 C; 131 G; 150 T; 0 other; gggggtcgaa atatggcgtc ttcaaatgtt gctcagaact tcacagctga aagcatcgtt 60 ccagaatcgc tggatgcagc acctgatgat caggttacgg tggtttatcc aggcaataaa 120 accgttctct ttaataaatt aacaccagct gaggtaagac cgcaacctga agtctccttt 180 aacgctgacc cgagtcaatt gtacaccttg gcgatgattg atcctgatgc tcccagtaga 240 gctactccaa ctttcaggga aattcttcat tggctcgtcg tcaatgtgaa aggggacgat 300 ctttcaaccg gacaaacaat tgccacctat agaggatccg gagcgccgaa aggcactgga 360 tcacacaggt atttcttcgt tgtttttcat caacctgggc caattgctgt agctggaaac 420 gatttggaag caaatagaag aaacttctcc attaggcaat tcgccctaga acatcaactg 480 ggcaacccta ttgctggaaa cttttttcag gccgaatggg acccgtcagt tcctgaacga 540 cgatgagtag cttaataaat tgcttggatt gataaaacaa aaaaaaaaaa aaaaa 595 // ID GT422712; SV 1; linear; mRNA; EST; INV; 843 BP. XX AC GT422712; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-843 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 641c5ef11e33caa29671ecfc3befbce7. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 19. XX FH Key Location/Qualifiers FH FT source 1..843 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 843 BP; 297 A; 162 C; 212 G; 172 T; 0 other; tataaataca atcaatttga atataacaaa ttcctgttta actaaggaaa attgggaatt 60 tttctcaaat taacacgcct gtctatcaac tggtatcgtg ttttgtgtgg tgctttattg 120 aaaaatgacg actgctgccc gaccaacttt cgagccagcg cgtgggggca caggccgaaa 180 tgagaaggat ctaagcgcca tttcacgcca gtactccagc cgcgatttac caggacatac 240 taaactgaaa tatcgggagc atggacaagg gactcaagag gaaacccgat caagagattt 300 ccgcagggag ctggaggata gagaaggcca taagaaagta ggcggatcca gaagcgccga 360 ccatgcaagc agtaaacggt tgaaactgga tcaggtgccg gctgcaagtc tggatgcaga 420 tgatccagtg gatccggaag acgaatctga tagcagtgag gatgaagacg acaccgaggc 480 cctgcttctg gagctgaaca acatcaagaa ggaaagagct ttggatcaaa ccaagaagga 540 aatcgaaaag cgtcaagaag aggagcggat cagaatggaa aacatcctca gtggaaaccc 600 gttgctcacg cattattcag caggagcggc caaagtggac cacaaagtca aaaagcgatg 660 ggatgacgat gttgtgttca aaaactgtgc taaaagcgag ccggaaaaac gcagcaatgt 720 gtttataaat gattcccttc gctcagagtt ccacaagaag tttatggaga aatatgtcaa 780 gtaaagactt gatggtttga gggtaacaaa taaaacattt actataaaaa aaaaaaaaaa 840 aaa 843 // ID GT422713; SV 1; linear; mRNA; EST; INV; 865 BP. XX AC GT422713; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-865 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ec3f9bd6e3321134ccd6e69ba9f74146. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 20. XX FH Key Location/Qualifiers FH FT source 1..865 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 865 BP; 299 A; 158 C; 156 G; 252 T; 0 other; gatttgatta tgctgaaagt gcaaaaaata aatttggtac tacataaaaa tgctccgttc 60 cccttagtta aaacctacag ttcttggtca aaatgagcaa taatttcagg aaagttttga 120 acgaatcttg ttcactattg gacacctata aaggaagaga taagattatt aggacattat 180 gctatctatc ccgtttattt ggagaactgc aatccaaccc ggaaatccaa cagaaatgca 240 atactttcag tacacaaatg tcggccacca gaaccacgtt aaggctcttg gacgatctac 300 ttgtattaag aagcacttta gattataaat tcggacagaa tgaagcagat aaatatatgg 360 cagtgatggt ggtcatgtcc aatattacag atcacatata tctttccctt gaaaagtttt 420 catggcttgc caagcataag ctattaactg gaattgataa tacaaaatgg gacactgcca 480 gttcagcatg ctgggtgttc acttcatacg tatctatttt aaaaaacgtc agatttcttt 540 tcatgatgga gagccacaag agctgcctcg gccaagttaa aaacatatca gacgaaaagt 600 tgagactact caaatggttt catatctgga ccgtgtgtag gtccctcctg gacttcacgc 660 atgcagttaa cactctacca ccaggatttt tatggtcttc aaaactgagt tctcgcttca 720 taagtctgat cgggtcaagc tcctcactga ttggactgta tttaattatt tacaagaaat 780 gcttgacgta attacgaata atatcgggtt ctagacaagt atttataaaa taaaatattg 840 taaaaggaaa aaaaaaaaaa aaaaa 865 // ID GT422714; SV 1; linear; mRNA; EST; INV; 842 BP. XX AC GT422714; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-842 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0ca923352d71fb53b0337a438b18447d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 21. XX FH Key Location/Qualifiers FH FT source 1..842 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 842 BP; 317 A; 130 C; 165 G; 230 T; 0 other; tgtcagtgtt tattagggag cagcgaaaga tccaggtcaa gatcaccttt aggggacgca 60 cagtttcatc caaagataaa ccacaaaata taggccttgc agcaaagaac cgactgaaaa 120 atatgagcat tcaaatgaaa ctcaattcca gcaagcctgc tccggctaat aaaccaaaac 180 tcagtgtagc tgctgcgttt agtcaagaca gcgatgacga acctgaagaa atgcccccag 240 aagccaaaat gagaatgcga aatattggga gaaacacacc aacttcatct ggaccaaatt 300 catttggcaa aacaaagcat ggattctcag atgctaaaaa gatattcgaa aagaacttaa 360 gggaggcgat ggaagctgca agagctgatg attaaatagt taagactagt taaattaatt 420 ttttattgtc aagtagagct ttaagcggca cagtagttca aacgcttacg tatatttgtg 480 attattgccg tagattggca cgtagttatt gatggggtgc cttctaatca acggaactgt 540 ttagtttgct gtttttagca aattaactta cgttttgtga ttgcattacc tttatcgatt 600 ttacgaagaa aattaatttt ccaattattg accaaatggg aagaatggga ataattgtat 660 ctatagtaga acttcaaacg tacaagtatt taataataca caatcaaaag aaagaataca 720 tttaatttga acgtaaaatg tttgttagta tagccgtcga accctaagaa atatgtatgt 780 atagaaagta ggtaaatgtt attgagaaat aataaaatgt tagtgaaaaa aaaaaaaaaa 840 aa 842 // ID GT422715; SV 1; linear; mRNA; EST; INV; 794 BP. XX AC GT422715; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-794 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 476ceba57352310d2c2e086e9bd7b503. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 22. XX FH Key Location/Qualifiers FH FT source 1..794 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 794 BP; 273 A; 140 C; 155 G; 226 T; 0 other; acgggctttt gccaataatg ttattctatc tggtggtaag cttatgacaa aaacgatggc 60 aagggactaa caaggaaagt tgccagccgt ttaactcgct atggagtctg aaaactagct 120 agcaggtatt gagggtgtgt aaacatattc aaagagagga gaaactactc tgcttaaatg 180 taatataatt ttacttttcc aatatagtat acacactaaa taaccgctag attcgtttaa 240 gtccatccgg tgtgttcaga cctacttgct tcctattcta ctttttagtt aactatttct 300 ttattcagga agcaccctgt tcaacgggat agaagaccgg ctgaaacagg aattggaaac 360 tagagtaccc cccaccatga aagtaaacgt aatcgctagc aaagaccgaa acattttcgt 420 gtggcgaggt ggttctattt tgggttcact cagcaccttt gtaaaaaaca tatgcatatc 480 tcgcacggag tatgaggaat ttggcatttc ggtaattcga aaaaaatgtg caaatcaatt 540 ttagaagaaa tgctattaat tcccagccat gcacaagttt ctttgcaacg gacatttggc 600 caatttagca ccatggttga agttaataag cgtaatttgg cacaaatggg ctagtcattg 660 ttagtaaata cacgtacaaa gaggtgaatt agtcatttac gtttatatta tgtaataggt 720 acaagatcct attggaacaa tgtgggacct aacaaatcaa cagtcaaatc aaaactaaaa 780 aaaaaaaaaa aaaa 794 // ID GT422716; SV 1; linear; mRNA; EST; INV; 887 BP. XX AC GT422716; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-887 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 318876b7e3024f326c685e41c7585266. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 23. XX FH Key Location/Qualifiers FH FT source 1..887 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 887 BP; 242 A; 214 C; 227 G; 204 T; 0 other; agcacatcat cggcaaagtg ctgaaagcct ggtttgactc tggaaaacta aagcgacagg 60 aagttttcat cacgaccaaa ctgcctcctg ctggtctgca tcccgatagg ctggaagagt 120 atctgctgaa atctctcgaa ctgcttcaag tggactacgt ggacttgtac ttgattcatt 180 tgccgattca tatacagtct tttggtggtg ccagcacaaa tctggcggct tcgcagcccc 240 aaccgaccga tcatcttgca gtgtggaaga aactggaaga gcaagtggat ttgggcagaa 300 cccgcgccat tggagtgtcg aacttcaacc aaagccaagt gaagcgcgtg tttgaaaatg 360 cgcgaatcaa acccgcagcc aaccaggtgg agttgcacat ttacctccag cagcctgagc 420 tggtcgatta cttgcaagcc aatggcatcg tgcctgtatc gtacttttcg ctgggaaatc 480 ccggaattaa cgaatttctg gtgaagaacg ggcgcccgcc aaggaacgtc ggcagtgagc 540 tgctggatga tccagttgtg cgggaaatcg ccactaaaca tgggaaaaca tctgggcaag 600 tcttgctgaa gtatctggtt cagaagaata ttgcggtgat cccgaaaagt gtgactccca 660 gcagaatccg cgagaatatt gggctgttcg attttcagct ggatgctgct gatgttggag 720 ctctgcaggc tctggataag ggcgaggccg gcagacgctc gaccatggct ttcaatcaag 780 ctatcgtcga tcatccagag tacccatttc cccgcattgc caggactgtt taaacccgtt 840 aattgacgca ataaacgcac actaattacg aaaaaaaaaa aaaaaaa 887 // ID GT422717; SV 1; linear; mRNA; EST; INV; 729 BP. XX AC GT422717; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_O24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_O24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-729 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 2005ad4c0c4eb87c1e45758c7086be15. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: O column: 24. XX FH Key Location/Qualifiers FH FT source 1..729 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_O24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 729 BP; 264 A; 120 C; 118 G; 227 T; 0 other; tcacgtcata gaatgcaagg gatcagttaa accaaagcat gtattgccat tgaagagcct 60 ccaattcgta gtcacccatg agaacgaccg attggtactg gtgaagattc cagaagatat 120 gctgaagaag gataagggtg atttaatatt ggaagtgcct caacttattg aagctatcac 180 aatgattatc gacgtgacca aagactcttc acttctgacg attctggata cgacaacgat 240 tgagcaccac atgaatggta aaaacggtac catcgaaatc cagcttggat tgcctgagcg 300 aattcacaaa gataaaagtg gccatctact tatagtgtca gccccttaat tcttggtcag 360 agcaagtagt ttatcatcgt aataaccaac ttctggagct gctacttaat ttaattgcca 420 aaactaatac atgagcaacc acttctaatc aaatgatttt tcataaattt cctgcctaat 480 gagctttgcg atagaaatat aaattattct ttgatattaa caggagaatt ataaatttga 540 aacatattta tataaataaa taaatgttga tcaaaaatca attaactatt ttatcatgaa 600 aaatatgttt acaatggaga catcatttaa ttttaaattt tgtcttgcgt ttattgttgt 660 ctatttgttg ttaacttagt acctataaaa taaacaattt tattttggcc taaaaaaaaa 720 aaaaaaaaa 729 // ID GT422718; SV 1; linear; mRNA; EST; INV; 817 BP. XX AC GT422718; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P01 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-817 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; cec99cc97fde6c31caaf47c0f56041b1. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 1. XX FH Key Location/Qualifiers FH FT source 1..817 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 817 BP; 253 A; 171 C; 170 G; 223 T; 0 other; cacatgttca gaggcaaact ttattgaaaa agctggcgct caaagatacg cagctgaaca 60 taaagtaatc atcgtcaatc cagacacttc acctagaggc ttgaacattc caggcgactc 120 tgactcctgg gattttggag ttgccgcagg cttctactta aacgccacgc aagatccgtg 180 gaaaaacaac tacaaaatgt atgattatat tatcaaggaa ctcttggact tagtgaactc 240 gaatttcaac actcaagctg gaaaccagtc aataatgggc cacagtatgg gcggtcatgg 300 agcgttggta ctttccctta aaaatcctgg gctgtttaag tcagtgtcgg cgtttgcccc 360 gatttgcaac ccttctattt gcccttgggg tattaaggcg tttagtggat atctgggccc 420 gaaggaaact ggccaatggg aggagtggga tgccaccgaa ttggtcaaga aatacaaagg 480 gcctcctttg gagctgttca ttgatcaggg actgaatgac aatttttatc ctgcccaact 540 aaatcccaaa aacctagtgg aggcttccaa aggcgcgggg gtgccggtaa tctttaaaga 600 aagagaatct tacaatcaca gctatttcta catagcgtca tttattgggg agcatatcgg 660 ctatcatgcc cagcatttga aataactctc atgcacttga atccttctta cgcttgcact 720 gtttgatttt aaagtaaata tttaatatca ataattataa acaatattct ttacacacaa 780 taaatattgt tgcttgttcc aaaaaaaaaa aaaaaaa 817 // ID GT422719; SV 1; linear; mRNA; EST; INV; 426 BP. XX AC GT422719; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P02 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-426 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 29023cf12bd04ae6b07f18df94c4222f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 2. XX FH Key Location/Qualifiers FH FT source 1..426 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 426 BP; 133 A; 104 C; 91 G; 98 T; 0 other; ggggattcat ttgcagaaat aagcccgagc atcagcaaga tgatgatttc gaacaaagtt 60 tgcgtcctca gcttcgtgct gcttctttcc ctagcactca gctctagggc tagcaacatc 120 cacagattca atgcagccag acatccccta gatgtttccg gcctggaggc caatggtgca 180 cctgtaactg caattgctga caacgatgct actgaaactg caacagctgg tcctactgca 240 actgcaactg caacagttgg tcctactgca actgaaactg caacagctgg tcccactgcc 300 actgaaactg ctcatgctga tttagctgtg acctcaaaag gggttgtaaa aatgtacatg 360 gaataagagc tgcaattaaa taatcgagtg gtttgccacc tggaaaaaca aaaaaaaaaa 420 aaaaaa 426 // ID GT422720; SV 1; linear; mRNA; EST; INV; 794 BP. XX AC GT422720; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P03 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-794 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 569e1266cb9152087f3ef55c8b273186. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 3. XX FH Key Location/Qualifiers FH FT source 1..794 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 794 BP; 223 A; 168 C; 180 G; 223 T; 0 other; gggggtcatt ttgtgttgca tactgaggta tccaaaaaca tgaagagtgt cgctgttgca 60 gttatatgcg ctttggctgt aggctttgca tccgctaaac ttagtggatc tggtacaacc 120 acccgttact gggactgctg caaaccatcc tgctcatgga aagagaacag tggaactcat 180 gatgctgttt actcttgtgc agttgatggt gaaactaaac tgaatgcttc cgtgcattct 240 ggatgtgact ccgatggtac ttcttacatg tgtaacgacc tacaaccctg ggccgtcaat 300 gacagttttg cttatggttt cgtagctgct tcattcgctg gtggtgttga cgttcaatac 360 tgcagcgttt gtcttaagct taccttcacc aacgctctat cgggcaaaac ttttgtagtt 420 caaaacgtta acactggtgg tgacttggga gctaaccaat ttgacattca aatccccgga 480 ggtggtgtcg gtattttcac cagaggttgt tcaactcagt ggggtgccga cgaatccggt 540 tggggtcagc aatatggtgg agtatcttcc gatgcagaat gcagcgaact tcctgaagtt 600 cttcaacctg gttgccactt cagattcggc tggtatgaaa atgccgacaa cccacaagtc 660 aatttcgaac aagtcgaatg ccctgaagaa cttaccatca aaactggatc cgaaaataac 720 gatttgacca tgaccctaaa ttaattgaca tttaaaaaaa aataaatctg taaatacaaa 780 aaaaaaaaaa aaaa 794 // ID GT422721; SV 1; linear; mRNA; EST; INV; 846 BP. XX AC GT422721; XX DT 02-JAN-2012 (Rel. 111, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P04 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-846 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6cbf9e58e963884e96fab4730471f1de. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 4. XX FH Key Location/Qualifiers FH FT source 1..846 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 846 BP; 244 A; 205 C; 198 G; 199 T; 0 other; gccaattggg ttggtcattg agccacaggc gaaaaaaaac gcatttttga agaagaggcc 60 catcgacatt attactacag gggagtataa caaagtgccg atgatcttcg gctacaacaa 120 caaagaggga atccttttca actacagacg ccatctaact ggtggggttg agaaagtgtt 180 gccggaatac ttcattcccc ataacgtgaa tctgaggtcc aattcgaccc tgcgctggac 240 ctacgtagag aagctgcaaa acctctactt aaacggcccg gacaaagacg acaatgctgt 300 tatgattata tcagacgctt ggttcattag cggaatcgtg gggagcgtga agaaccactc 360 gcaaacttcc ccatatcccg tctacttgta caagctgtct ctggacaccg agctgaattt 420 cgtgaaaaaa gcctgccagc taactgagat tgctggctgt tcgcacggcg atgacctcgg 480 ttacttcttc aaaacgccat tcacgcccca aattgcgcca tccagcatcg aagatgcgtc 540 tttaaggcgg cttgttcggc tttgggccaa ctttgccaga tcgggcaatc caactccgga 600 tgccagcgag ttcgggctga gctggaagcc tgcgatgacg cgtcaattga acactttgca 660 tatttccaaa gagttgacga tcgaggttaa tccggaagga aaacgaatgg atttgtggcg 720 gcagctctat catcaaagca atgccactgc taaatattta taacttccat cctccggttg 780 cagttgaatg ttgcctcaca gttgctggca taaacgcatg aagttcccaa aaaaaaaaaa 840 aaaaaa 846 // ID GT422722; SV 1; linear; mRNA; EST; INV; 793 BP. XX AC GT422722; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P05 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-793 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c796595b2ee4001ba83067d8d21863d5. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 5. XX FH Key Location/Qualifiers FH FT source 1..793 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 793 BP; 263 A; 145 C; 154 G; 231 T; 0 other; tcgcttgcta agtttcacgc atctggaatt gggctcagac atttgcatcc agatgtcttc 60 gagtccaagc tgaaacccca tttttactgc ttcactccct atggtttaga tgaattggta 120 ttagctagct taattcctgt tctggaaaaa agcgacaaat ttagtaaagc agaagtggac 180 gtggttaaat cagtattcgc acagaacgtt aaaaaactgg acgttagtga agcttggggc 240 acgatcatcc attacgattc ttgggtcaat aatattctct acaaactaga aggaacgaaa 300 ccagttcaca gcattttagt cgacttccaa gtcttcgact acggcactcc attaaacgac 360 gttatgtttt tcttgttctc cagcataaag caagatttgt ttgaatcgca aatcgacaac 420 ttgctcaatt attactatca agggctcgta agcaacttaa agacattggg ggtggatatt 480 tcaaagtttt cccgctccag ctttgaccaa gaagtaaaaa tggcagtgca aaattatgaa 540 tattatcatg caatgtggat gcttcaaccc ataatgcacg aagcatttaa gtctgttgca 600 agtttaaagg acggaactga agatcctaca agttggtcaa cggagcacca aaataaagtg 660 aaggatataa ctcgcatgtg cttcgataaa ggatggctgt gattattacg ttagtctgga 720 ttaaaatggt tttaatgtaa attgtatagt ttaaaaaata taccacttta aatatccaaa 780 aaaaaaaaaa aaa 793 // ID GT422723; SV 1; linear; mRNA; EST; INV; 798 BP. XX AC GT422723; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P06 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-798 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f926a2693415faddee558256b5546fd9. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 6. XX FH Key Location/Qualifiers FH FT source 1..798 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 798 BP; 235 A; 210 C; 183 G; 170 T; 0 other; acagcagtgc tttgcgtgca ttggcagtcg aatataaggg gttgcaggaa gagccagtgg 60 agggttttcg cgtcaagctc gtcaacgacg acaatctctt cgaatgggag gtcgctattt 120 tcggcccccc cgacactctc tacatgggag gctgctttaa ggcaagagtg aagttcccgg 180 ccgactaccc ctattctcca ccctccatca ggttcttgac caaagtctgg cacccgaacg 240 tctatgaaaa tggcgatttg tgcatcagca ttttgcatcc gccaatcgac gatccgcaat 300 caggcgaatt gccctgcgaa cgttggaacc ccactcaaaa cgtcagaacg atccttctgt 360 cagtcatttc cctgcttaat gaaccgaaca cgttcagtcc tgccaacgtg gacgcttctg 420 taatgtaccg tcgttggcgc gactccaaag ggcgcgacaa agaatacgag aacattatca 480 ggaagcaagc aatggcagcc agagctgaag cggaacgtga cgggatccaa gtgccactca 540 cactcgaaga ttactgcata aaaactcaag ttaaacccag caactccgag tctcaagtag 600 aaatgacgga tttctacgac gatgactacg acgaggccta cgacgatctg tcagacaatt 660 cggaatatgg cgatgaagac gacaatgaca gcggtaacgg cgaatcgtga taacgcatgc 720 acatccagcc gattgccaca ttctatgtac atttaaattc aatacaagcc actcttttcc 780 aaaaaaaaaa aaaaaaaa 798 // ID GT422724; SV 1; linear; mRNA; EST; INV; 814 BP. XX AC GT422724; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P07 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-814 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 08115c23c5683994e686263f2a5c50f1. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 7. XX FH Key Location/Qualifiers FH FT source 1..814 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 814 BP; 234 A; 150 C; 144 G; 286 T; 0 other; gtcaaaaggg atccctcatt aatatcatcg caaagtgata taaactctgc ctaccataaa 60 atccatcggt attttaggag attgtttaaa tttgaacaga tggactttca attcgcattg 120 tggcaaatgt tttatctgtt agcagctccc cagaagctga cgaaagtgtt ccgggcgcga 180 aaaaatttca agtctcaata cgcacgtgac gatccagctt tcttaatact tttcgctggt 240 gctctgtgct tgacctcgat tggttttgcg ctcactctcg attatagctt tcttcaattt 300 accaaattcc ttgtcataga aatttttatc gattgtatag gaataggtgt tgttattgca 360 acggcgctat ggtacatcac caataagttt ttaaagccga aaaatatgcc tcaagatgtc 420 gagtggggtt ttgctttcga tattcaccta aacgcatttt tcccaccttt gattttgttg 480 cactttttct tattaatttt ctataatttg tttatgcaag agagcggcac gagccaacca 540 tttgccgtgt tctatggaaa tgcgttttgg ttagcagcgg ctgtttatta tgtatacatt 600 accttcctag gttataattc catgcagata ttaaaacata ttaatttatt tttaatacct 660 attcctaccc tgatttgtat atacttgcta actctgtttc gcggcattaa tatttctata 720 agcgtctttg acatgtacag gcagagaatt gtgtaagact ttgttaagct aaaagtaata 780 aaatatgcta ctggtgcaaa aaaaaaaaaa aaaa 814 // ID GT422725; SV 1; linear; mRNA; EST; INV; 483 BP. XX AC GT422725; XX DT 18-NOV-2009 (Rel. 102, Created) DT 28-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P09 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-483 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d057822a2a307b9723ede52e028dd1b5. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 9. XX FH Key Location/Qualifiers FH FT source 1..483 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 483 BP; 156 A; 113 C; 117 G; 97 T; 0 other; ggggggctag aagcagacag acaagttcac ataaccacaa acctatgttg tagcacttgg 60 tgaaaattta cctaaatagg ccagtttcaa tgttttccgc ggtgcgttcg ttactaagcc 120 gctccagagg cctgctgccg agctccagcc ccaaaatggc cttccttgcc cccaacagtc 180 ccagcctgac catcagctgt ggcatgaaag tagtggggct actcagaaaa aggtgcaaag 240 actgttactt cgtctgcagg caagagcggc tgtatgtaat gtgcaagacc cacggaagac 300 acaagcagat gtcaatggtg aagaagccga aaaatacctg gaccctaacg catgccaccc 360 agagcaagca caggccgtgg tagagcgcat tttgccagtt tgtataagaa aatcaccgaa 420 tagtggacag cgattagttt gcgataaata tagattgtaa caagcgaaaa aaaaaaaaaa 480 aaa 483 // ID GT422726; SV 1; linear; mRNA; EST; INV; 793 BP. XX AC GT422726; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P10 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-793 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 3c7b0c6fa9190a9fcfa58b815d427825. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 10. XX FH Key Location/Qualifiers FH FT source 1..793 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 793 BP; 218 A; 212 C; 189 G; 174 T; 0 other; caactcctgg aagggccgtt atgatacacc cgacagcaac ctcaccgccg acaccttcca 60 tggcttgttc tacgactggg agcctgtata ccacgaccca ccttctaaca gatggtttgg 120 aatgcagccc tggtcgaccg atcgtctggc ccagtactac tatgagacca aagactccaa 180 ggcggaagcc atcctgaaaa agtgggttgc gtgggtgatc tcagccatca agctggagaa 240 cggcgacttc caaatgcctg atcatctaac ctggtcggga aatcctccag acatccatgt 300 gagcatcgga gtttacacca aggaaatcgg aaccgcttca gctactgcga ggactttggc 360 ctactatgcg gctggttctg gggatgagga agcaaagaca gtagccaagg gcctcttgga 420 cgccttgtac aaatacgcaa cagacaaggg aattaccatt ccagaaaccc ccgatcagta 480 cggaaggttt gaggagcccg tctacgtgcc atcaggatgg actggaacat accccaatgg 540 cgatgtcatt gactcttcag ctactttcat aggcattcgc tcctggttca agaatgacgc 600 ccaatggtcc aaaatcgagg ccatcttgaa tggaggtgaa atccccgagt ttgagttcca 660 cagattctgg gcccaaaccg atgttgccat tgccttcggc acctacggca tcctgttcga 720 cgagtgaacc agtagtgatt ttttcgtaat ttgaccaaat aaaccctcgt acttctaaaa 780 aaaaaaaaaa aaa 793 // ID GT422727; SV 1; linear; mRNA; EST; INV; 801 BP. XX AC GT422727; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P11 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-801 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 04c79d1fa5ca4c643de5555a070c9630. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 11. XX FH Key Location/Qualifiers FH FT source 1..801 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 801 BP; 279 A; 151 C; 171 G; 200 T; 0 other; taagggactg cccgcagttc ccgaatcagt tctgaaacac cgaaaaaggc gcgcagcttc 60 aagagctaaa cgagtacagt cggacatcaa gaaaaagtcc gatcaaatta aaaatcgtaa 120 agaaatcttc aaacgtgctg agcagtacgt aaaggagtac agattgaaag agcgcgatga 180 aattcgtctt attcggcaat ccaaaaacaa ggggaacttt tatgttcctg ccgatgctaa 240 gctggctttt gtcatcagaa tcaagggaat caacaaagtt gcccccaaag tacgaaaagt 300 ccttcagttg ttcaggctcc tgcaaattaa taatggcgtt tttgtgaaac ttaacaaggc 360 tacaattaat atgttaagaa tttgcgaacc ctacattgcc tggggctacc caaacctcaa 420 gtcagttagg gaactgattt acaagagagg atttgccaag gtgaatggtc aacggattcc 480 tattagtagc aatcaaattg tggaagaccg attgggcaaa gctggcctaa tctgtgttcc 540 tgatttaatt catgaaatat ttactgttgg acccaacttc aaatatgcat caaacttcct 600 gtggccgttt aagctaaata ccccaactgg aggatggcgc aagaaggcca atcattatgt 660 tgaaggtggt gactttggaa acagagaaga caaaatcaac gaacttctcc gaagaatggt 720 ataaggagta gttttttatt ggaaaaaaga aacgttcgtt tctctaataa atgaactgtg 780 ttccaaaaaa aaaaaaaaaa a 801 // ID GT422728; SV 1; linear; mRNA; EST; INV; 799 BP. XX AC GT422728; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P12 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-799 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 82b2b80624788c4011db68b271e96802. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 12. XX FH Key Location/Qualifiers FH FT source 1..799 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 799 BP; 258 A; 158 C; 184 G; 199 T; 0 other; cagcatgaag gttccaggca ggagtacctg gaacatatct tcaaagagca caatttacat 60 ctggggaatc actacaactt agtgtttatt gatgaactaa ttgaacatgt ggactcccaa 120 ttgcaagctt tacgatgtct gttttgcaag ggcgtgtttc cggagcgggc agtgctcaag 180 gagcacatgc ggaagaaaat gcacaagcaa atcagccctg aggatgtgga atatgataag 240 ttctacataa taaactattt ggaagttgat aagagctgga gagtcctgca gcatgaaacc 300 gacaattacg ctttggaggt cgactatgag gagaactgcg atcaggagta ttcggattgg 360 accgaagctt ctggccagat caccttcctt tattgcgcgc actgtgaaac tgatatgggc 420 gccattttcg accacataaa aacgcaacat aactttgatt ttgcacgagc caccaaagag 480 ttggactttt atcagaaaat caagctgatc aactacacca ggcgaatgat ttataggaag 540 acgtgcccgt attgtacgga agagttgggc gattctgcaa agctccaagc acacttagca 600 caggagggcc attgtaaagt gccctcagac caggttttcg atcaagcaga atattacttt 660 tccacctatg aaaacgacac gtttttgtat ctaatcgatg atattgacga ttgaaagctg 720 gcaataatat ctgttttacc tgaagcgtta taggaatata aagcggctta gtcgcaaacg 780 caaaaaaaaa aaaaaaaaa 799 // ID GT422729; SV 1; linear; mRNA; EST; INV; 858 BP. XX AC GT422729; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P13 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-858 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; f0c385acc56e69afce754c7ee274ac68. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 13. XX FH Key Location/Qualifiers FH FT source 1..858 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 858 BP; 318 A; 129 C; 146 G; 265 T; 0 other; aagtagcgga cattaaatca aaactgacga actgtggaca accaggccat catggactgg 60 ggaatgtgaa aaataccagc acagttaata gattgaccga tgcctcaaag ttcacaggaa 120 cgcacaaaca gagatttgat gaagccggaa aaggcaaagg cattgctgga cgaaaagacc 180 ttgttgattc atccggttat gtcagtgggt atcaaaacaa agacagctat gagaaaaaag 240 aataaaagaa aaaccatatt tattaaaaaa tatatgctta agtttacacc gtgcactata 300 ctcgagaaat gagtaactta tattcaaata tttgtactcg tgtctgcttc caatggagct 360 tccagttttc ctttgcacaa gagtttgttt aaatagaacg tttttgatat gtcaagtact 420 tatttagata attataagcg gttgtactcg tttagagatt tttgcgtagt tttagtttta 480 tattttttca cgatcgttta aaaatgttaa actcactaga cacttacaat taaggtacgc 540 tgtgctggaa tgtgggacat tcttttaaat aaaactgcac acttcttcca gtctatttca 600 ttaaaagtct atgtatcata taatatgata gtaaatgtga tcgtacaaaa gtctagtgca 660 atgttgctct aactgaattg gtgtaaacta aatcaaaagc tatatatttt aggtatacca 720 cgtctatttc gaactccaaa cttaattagc ataaggtaaa gaataatatt tttactttcc 780 ttctcgtatg tagaatgtga aatttatatt aatttattaa aagacaataa aataaacaag 840 caaaaaaaaa aaaaaaaa 858 // ID GT422730; SV 1; linear; mRNA; EST; INV; 744 BP. XX AC GT422730; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P14 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-744 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 16d77b1ed08c925ee6bbde7447f39842. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 14. XX FH Key Location/Qualifiers FH FT source 1..744 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 744 BP; 258 A; 154 C; 165 G; 167 T; 0 other; ataagacttg caacaacgag attatcaaga tgcaaaggct caatcctgat cagctacaaa 60 acatgaccgg actggaatat atgctgttgc atgtccaaga gccgattctc tatgtgatta 120 gaaaacagca ccgccacagc ccccaacagg tgacgcctct ggctgattac tacataatcg 180 caggagttgt ctaccaggcc cccgacttaa acagtgtgct caattcaagg ctgctgtcgg 240 ctgttcatca tttacagtcg tcttttgaag aagcgtcagc attctccaga taccacccga 300 gcaaaggcta ctactgggac tttaagtcgc aaaagggccc tgaaaaggta aaaatcccgg 360 aaaaagagaa gccgaaagag gaatccagct cgttgtttca acgacaaaga gtggacatgt 420 tactggggga gttaattatg aaatttccgc tgccaacgcc accgcagccc aaaacggcgg 480 ccactgctcg agtgaaaact gaaagtgaaa atggagcgca agatgccaaa gacaaggctg 540 atgttaaaca ggagactaag gtggtgaaac ctccagcgga caaaaagccc agactgaatt 600 aactggattt gtgcatttat attgtattta cactttattg ggtggttttg gtggccagca 660 taatttaaaa tttatagaag taaataacga aatgtaacag aaagttgata atacataaat 720 taattgaaaa aaaaaaaaaa aaaa 744 // ID GT422731; SV 1; linear; mRNA; EST; INV; 780 BP. XX AC GT422731; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P15 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-780 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e1706a20fa5a65892433cb1c5ed64e52. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 15. XX FH Key Location/Qualifiers FH FT source 1..780 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 780 BP; 279 A; 132 C; 107 G; 262 T; 0 other; cagtacagca ttgaaaattg caatggaagc aggacatcgg cacataggcg tgcttctata 60 cgcgcatgaa agaacatttc aaaaatctca acccttgaaa ctgaaaaaga ccaattctct 120 cagtccaaaa acccccatcc tcaccattac ctattaggag tagtcatcgt gccttaactg 180 atacaaaaaa ataaatttta gaactgattt tcaaactacc cacgccaccg atttgtttat 240 ctcaacatgc tcatcttcga agtatatttt ctgaataata caccatacca ttttctagca 300 agtctgctca atggttctac ataaaacttg gacaatatct caaatgggag taatgcgtca 360 atttgttgag aaatggtttg cttagacaaa tttaattgtg tcatatacct acattttgtt 420 acttaactat tgttaaatag tataataagg agtctgcagt ttaaaaaata tttactagtc 480 taagcgaaaa gaaaacgaaa tctaggtagt tttgtatata tttaccagtt atacacccta 540 atcgttgtta cataattttt actagaaaag ttgtaattct atcacaagtt tatttaacaa 600 actaattttt atgtgaaggt tttgatagaa acactgtctc aattatataa gtcccgaatt 660 gatgatgaat ttagtgttaa cattccagta tacttacttt tacatgtttt tcttgttgta 720 acatattgta ttttttgtca ataatatata ttttagacag tcaaaaaaaa aaaaaaaaaa 780 // ID GT422732; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422732; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P16 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 20fd187f289f9dcf61452eb46c887c82. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 16. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 272 A; 97 C; 147 G; 300 T; 0 other; gtacgttctg atatcgagga agcacgggac aaaaagctga ttttagtaat gttcatgctt 60 tagttttaaa aaaatttaaa gttttaattt tgtaaataat ttgtaaaatt cctattaatt 120 atataaatat cttaagttgt ttgcccaaat cagggggcgc gatataaaaa cctctcttta 180 gaaagaagga gcgttttcga agatcggttg ctttttcgtt atgaagctat gtagacttta 240 aataggtttt tcgtatacat cgggtatgat caattaaggg cgaggaacaa aattgtggta 300 acaattgaac tatctgaaat gtgacgaaaa tatttttcta aagctcttta agttaagcgt 360 taagccttat gtatgtttgt aatacttttt atattgattt tctaacgcag tggatttgca 420 gttttgctta ttaagttcat aattactgtt tgttgctttt cgttacaggg aactcgtgca 480 taataattag gcaagcattc aataccctaa tctagtcgca atcgagaaca tattgatgtt 540 atattttttt aattgaataa tctcgtcttg tggaaattaa gttgcaatag tttgaactat 600 caggaagtac aatcgaattt tagttcagat gtaaatatct gtgaaatgtc ttcttgattt 660 agtaattaaa tttgttcatt atagatttta gccatatatt taggaatgga tgtaggattt 720 ctattgctgt ctggcgaagg taatacgaat aagatttatg cttattattc gcttaaataa 780 aatcacggag aaatgctcaa aaaaaaaaaa aaaaaa 816 // ID GT422733; SV 1; linear; mRNA; EST; INV; 822 BP. XX AC GT422733; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P18 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-822 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; b3f33a7f7b9867d063a69bc28826ef28. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 18. XX FH Key Location/Qualifiers FH FT source 1..822 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 822 BP; 234 A; 147 C; 162 G; 279 T; 0 other; gttggtgttt ggtattgtgg caatgtttgt gcacaacaag atcgtgcaac tggtgtatgc 60 ctccctaggg gctctgatct tttccatcta cttggtctac gacactcagc tgatgatggg 120 cggcaagcac aagtacagca tttctcccga agaatacgtc tttgccgctt tgaatctgta 180 tttggatatt gtcaacatct tcatgtacat tttggccatt attggccatg cccgggatta 240 atcgtttcat ctttcgttgt ttcttgtttt attttcagct ctacaatgat tcttgtataa 300 atattgttta gtagaccaca ggagcagttc gggttaaggg aacaacgcgg attggagaat 360 ttcgtttgac taattattct gctgagagcg tctccaagat atcgttttat tgctgaaagc 420 aaagcgcctc gaaaactatc aataatcagc atttgtttag gttacttgaa caagtctggc 480 ctgtactgaa cgtccttgca aaagtttctc agtatattaa aaaagcgata atattacttt 540 tagtccttag attaaggggt ttccatactg tatgcagaaa ttccgtacgt atgtgataag 600 tttgtattta atgtattatt attaatccac ttaactacaa tgatcggttt ctagtgggat 660 gtgcatctta aaggtagctc tggcacaaac cacacacttg gaacgaacag ttctattgtt 720 aaccgcttag tctcatctag aattatatgt taattatgcc aggttacata cagtacatct 780 gctgtgaaaa tatatgttta ataaagaaaa aaaaaaaaaa aa 822 // ID GT422734; SV 1; linear; mRNA; EST; INV; 486 BP. XX AC GT422734; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P19 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-486 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 26418a8eddfeddb137db6ce2bfb140cf. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 19. XX FH Key Location/Qualifiers FH FT source 1..486 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 486 BP; 133 A; 110 C; 112 G; 131 T; 0 other; ggggaactta aacgaaggca gcaaaattcg tcatggtgcc taatcaactg atcccagctt 60 tattggttct tgtgctcgtg cttgcttcag ctgccgaggc tcagctgaga tgctaccaat 120 gtaactcgta ccgggacggc gctaggggat ctctgtgcca caatcccatg gctcccaatg 180 tttctacggt ggtttgcagc tccgacgaag tttgttccac agtttcctat ataactggac 240 aaatcctggt tacttccagg gcgtgcgatc caagatcaaa ctcttgtcgc aacatcgaaa 300 gcggacttcg ggcattatat ccagacctga gcaggttcta ttgttcaaca tgctcatcaa 360 acttttgcaa ttccgcagga aaagttggga tttcaatggt ccttgccata ggctgcctgt 420 taatgtccag ggtgatttga acaagggaaa atatatgatt tgatcagtaa aaaaaaaaaa 480 aaaaaa 486 // ID GT422735; SV 1; linear; mRNA; EST; INV; 791 BP. XX AC GT422735; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P20 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-791 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6cfd0bb4083a7697160e775819d3b088. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 20. XX FH Key Location/Qualifiers FH FT source 1..791 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 791 BP; 270 A; 148 C; 190 G; 183 T; 0 other; cgaaggaaaa gatgatagcg taacaagtgc agactatttc agccactgtg ccatgctcgg 60 catcataaca agggctttgc cccaagtgac gctaatttct gaggaaaacg gcattaattg 120 tgaaaaggaa tcggcagccc aaacgtacaa tcaaccatta aatctgcagt ttgattatcc 180 agcggcgtat gtagaggcaa gcgacgttac aatatggata gatcctttgg atgctaccgt 240 ggaatataca gaaaagttgt atcaatacgt gaccacgatg gtgtgtgtgg ccgtaaacgg 300 taagccgatt attggggtaa tccatcaacc gttcaatgag tcaacttctt gggctgtggt 360 gggttttgga cattcggcca atcttaacga gaaggtacat gggacggatt cgaacttgcc 420 aaaaatcatc atatcacggt cacacagtgg aactatagag caaacattga aaacaaagtt 480 taaggatttt aaactgatag ttgccgctgg cgcagggtac aaagccttaa gagtcgcaga 540 caatacggcg gatgcttact tgcacagcac ggcgatcaaa aagtgggata tttgcgctgg 600 aaatgctatt ctgagcgctt tgggcgggaa aatgactacc aaacacggac aaacactgac 660 gtattacaac gacactaatg tgatcaataa agagggctta gtggcaacgc tacgaaacca 720 tgaagcgttt atagataagc tgtaaaataa aatatatgga gtttatatag agcaaaaaaa 780 aaaaaaaaaa a 791 // ID GT422736; SV 1; linear; mRNA; EST; INV; 779 BP. XX AC GT422736; XX DT 02-JAN-2012 (Rel. 111, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P21 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-779 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; bee8a7cbad2a769cfbcaa64686fa81ec. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 21. XX FH Key Location/Qualifiers FH FT source 1..779 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 779 BP; 229 A; 155 C; 164 G; 231 T; 0 other; ggattcaatg cattgagaaa tcaatttcct tatcaagtat cagtgcaacg tctttggatc 60 ttaacatact ctcatgtttg cggtggcagc attatttcat ccaattgggt actcactgca 120 tctcactgca ccacatctgg aacatatcga gttgtagctg gcattgtctt gcaaactgac 180 gaaaatgttt ttggacaaca aactcttgca gtttcagaga ttattaatca tccactttat 240 cccgagactg atgctgtagc tccaaatgat atatcattac ttcgcctatc tagaagcctt 300 atttatgtag aaaatgttca accgattcct attccacacc gaggatatgt tgctgaagga 360 gacgcagttc ttaccggatg gggcttgaca cgacctagtg ctctgactat cccaaatcat 420 ctccaatttg cttatcttcc tatcgtccca ggagaagaat gcgacaacat tttgaacgac 480 ttgctcggag atcgtaatcc ctttgatgta gaactaaacg tgtgctctgg aatcagaaat 540 ggtggagagt cggcttgcag tggagacagt ggtggtccac ttgttaataa tggtttactt 600 gtcggagttg tgtcctgggt attagttcca tgtggtggac ataatgcccc aacagtgtat 660 ggcaaaacat ctgcatttac agactggata gtgcaaaata ctaacggaga agtagagcct 720 gcaattttgt aaaaattgtg tttaataaaa gcgtgacttg gaaaaaaaaa aaaaaaaaa 779 // ID GT422737; SV 1; linear; mRNA; EST; INV; 807 BP. XX AC GT422737; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P22 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-807 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; b2933ed411bd4bbad03d2b35fd46f148. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 22. XX FH Key Location/Qualifiers FH FT source 1..807 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 807 BP; 317 A; 116 C; 163 G; 211 T; 0 other; aaagacattt tctgcatctg atatataacg tgcaagtgaa gaagacatgt tttaatgtat 60 atgataattt ataaataatt ggaaccacct aatattctaa tcgtatatga aagcatcgtg 120 cagcggtacc aattttagcg gaattaagaa tcagtaataa attaaaatag tgaaaatgga 180 gaatcccaaa cacatcttgc tattcagtgg aaaacgtaaa tctggtaaag atttcatttg 240 tgagaagctt aaatccataa taggagaaga caactgttgt atagtacgca tatccggccc 300 attgaaacag ttattcgcac aaaatcataa tttgcaaata aatgaattaa tgagcgatgg 360 accgtacaaa gaacaacaca gactaaatat gataacttgg agcgacgaaa tcagaaagga 420 ggatccgggt tacttttgca gagcagctag caaatcagca ccaacaagaa gactgtggat 480 tgtaagcgat ataagacgaa agactgatat caaatggttt aaggagaatt acagtaattt 540 gaagctaatc agagtatcgg ctgatttaaa aatcagaaag gaaagaggat ggatattcac 600 aactggggtg gatgatgccg agtccgaatg cgacctagac gattttcact cttgggatat 660 ggaaatggca aataataatc acgaagatta tgaaaatgcc atccgtacaa ttttgaactt 720 aatttcctaa tattgttgga tgttttttaa acctttggtg gcggtacaat acagaataaa 780 gtgactgaga aaaaaaaaaa aaaaaaa 807 // ID GT422738; SV 1; linear; mRNA; EST; INV; 795 BP. XX AC GT422738; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P23 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P23 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-795 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e43a9f92aa59623057ddb89b68203bea. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 23. XX FH Key Location/Qualifiers FH FT source 1..795 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P23" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 795 BP; 289 A; 164 C; 212 G; 130 T; 0 other; caaggaagcc attgaaaagc aacgggaggg aagagtggaa ctgtacaaac taccggctgt 60 aaacaaacag ctggcgctga agattatgtc cgatcaagcc aatgccaaga agaagccaga 120 atctgcaaat ttgttgcagg acaaccgatt caaggccctg tttcataatc ctgactttga 180 ggtgcaaacc accgctgaag agtatcgact attaaacccg gtgctttcga acatggacaa 240 tggcaaggca actaaaaaaa agatgttcgg cgctgtccgc gaagtcctgc cagatgatgc 300 agaggagatt gaggaacagg acagttcaat agcttcttcg gacgaggcca gttccgatga 360 cgagcacacc tggtcaaaag aagtgaaaaa gcaacatcga ctgatacaaa aggagcaaag 420 ggagaaagac agggccgatg aaaggcagca gctggcagaa gaagacgcag caaaacctct 480 gggcgcagca cgtggcctga aaattatcag gccgaaagtg aacaaggctc cccttggcga 540 gagagtggca aacgaaaccg actcggttaa gtacaactca gtggggaaca gagaaatgac 600 gttttccacg cggaaacggc gcgacaatcg agtggaggaa accaacaaaa ggcacaaaga 660 ggagaggaag aaggtggtgc ggcccgcttt cggctttaaa gcgaaacaga agcccagatt 720 taacagaaga aagtcctaaa cttgtaaatt atgaaattaa acagtaattg ttgtgccaaa 780 aaaaaaaaaa aaaaa 795 // ID GT422739; SV 1; linear; mRNA; EST; INV; 851 BP. XX AC GT422739; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO114.CR_P24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO114_P24 3', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-851 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5e4308376e23b4dbdb353ea3741b5bd4. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO114 row: P column: 24. XX FH Key Location/Qualifiers FH FT source 1..851 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO114_P24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 851 BP; 303 A; 129 C; 148 G; 271 T; 0 other; gctgaagctg ctagaaactg atattcaaga gctgatacgg gaagccaccg aagccaggca 60 gaatgcttac tgtccttaca gcaaatttaa agtgggtgct gcagttctaa gcacagatgg 120 aacaattttc actggttgca acgtagaaaa tgctgcattt actagtggaa tttgtgcgga 180 aaaatgtgct tatggtaaag cagtgagtga gggcaaaatt aaatttcgag ctgttgcggt 240 aattgctcag caggagtcgt tttttaccac gccttgtgga tcttgcagac aatttatgag 300 tgaatttggg aatgtcgatg tctatgtttc caaaccgaac catcaggatg tattggtact 360 cactttagac aaattactgc catatcagtt tgaattgacc aatcaaacat ttatatagca 420 agtcatttaa cagattaaca gttataaaaa ttttatttga acagcttata tacagagtgc 480 ctaataaaca tgttaaagta caacagctta aaattctaaa tttacattat acaattcctt 540 tttaagtgaa tttacataac gcaactgcgc tcccagtaat ataaaaacaa cttataatta 600 ggcaaaagca tataaataaa ttatttaaac atactattta tattaatatt atattaatta 660 aacatactat tgtgttaata gtgatttgca aatctatggt gatttatagg gattagcctt 720 tttaaaatgt gacctctctt gcaaaagtaa tcaaattttt aagaatgaaa aatgctgaaa 780 ttttgtggca ggtctgttct tcaactcgta cttttgcata agagaaattt aattccaaaa 840 aaaaaaaaaa a 851 // ID GT422740; SV 1; linear; mRNA; EST; INV; 812 BP. XX AC GT422740; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A01 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-812 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0b36cc14c218a6df8a06879270c3034a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 1. XX FH Key Location/Qualifiers FH FT source 1..812 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 812 BP; 266 A; 166 C; 184 G; 196 T; 0 other; ggggaacaag agtaaggtta ggagagttga gaaaaacatg cgaaagttta ggaatttcca 60 aacaaacgcc ccgacaaatt aaggaaattg actgaaatgg ttgacgctcc gcaataccca 120 actatcaacc gagaggcacc gaatatacta gtaacgggaa ctcccggtgt tggaaagtcc 180 actctatgta agaaattggc cgaagtgact gcattgcgtt ggttggaaat ctctaaaatt 240 gccaaggata ataactgttt gcaggagtac gaccccgaat tcgatacaca tgtgctggat 300 gaggataagc taatggacgg cctggaggaa gttatgagca tagggggaaa catagtggac 360 tatcacagca gcgaattttt cccagagcga tggttcgatg tagtgttcgt tctcacggcc 420 gacactagca gtctgtatga tcgactcaat ttgcgtggct attcaggcaa aaaaactaga 480 aagcaacatc gaatgtgaaa tcttcaaaat atgtctagac gaagcaaagc aattttacag 540 aacccagata gtacattcac tggagagtat aaatgaagca cagctagagg aaaacgttca 600 gaaaatatgc caatggatac aaatgtggct caacaatcga gaaatgaatc cccaattaaa 660 cagttaggct ttattagact taagactcac cttagcctat tcataatgtc aactcccgcc 720 tgatgccgaa cgcaactggt cacataatcc aaaggaatct gctttttctg tttgcagtcc 780 cttttgatat cctgcactgt gtgctcgtcc ct 812 // ID GT422741; SV 1; linear; mRNA; EST; INV; 756 BP. XX AC GT422741; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A02 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-756 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; c9bd3af625ca21510891c7147f4c0bdc. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 2. XX FH Key Location/Qualifiers FH FT source 1..756 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 756 BP; 204 A; 194 C; 212 G; 146 T; 0 other; ggggagcgat gagtgtattt tttcctaaac gctgttacgt atagtaggta gaggggtaga 60 ggtcacgtcc tgcactgaaa aacacggaac atcccccctg aaagctcctg acaatgctga 120 agcctcgact tttcgcacat gttaagcccc aattgagatg caagtcccac ttggaagccc 180 tacgcgaaaa aatcgccaat gggcccaact tgcaggattt catcaacgac cccctgccag 240 cagaagagac caactcttgg caggagtacg agggcaagtt gaagagggaa aagggcgagg 300 agcaacgctt aaggctgccc ccatggctga aacgaaccat ccccatgggg aagaactaca 360 tgaaaatcaa agagcaactg cgaggcctca atctgcacac tgtgtgcgaa gaggccagat 420 gcccgaatat tggggaatgt tggggcggag gtgagcatgg gactcaaacc gccactatca 480 tgctgctggg cgacacttgc acgagaggct gcagattctg ttcagttaaa acagccaggg 540 ctccgccccc accggacccc aatgagccaa tcaacacggc caaagccatc gcgtcgtggg 600 gcctggacta catcgtcctc acgtctgttg accgagatga cttgcctgat gggggctccg 660 cccactttgc tgaaacggtt agggagatta aaaataggaa ttcgaatatc ctagtggagt 720 gcttggtgcc ggactttcgg ggcgatttga agcaag 756 // ID GT422742; SV 1; linear; mRNA; EST; INV; 803 BP. XX AC GT422742; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A03 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-803 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 9d07783fffb84d006af0c5c016a7043a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 3. XX FH Key Location/Qualifiers FH FT source 1..803 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A03" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 803 BP; 257 A; 146 C; 208 G; 192 T; 0 other; ggggagtgtg gcgggcgcgg ccatattggg aattcgtgag acaagatctg aattgtgagt 60 gtagtgtggt gaaaattaat ccgaaagcat cggaaaatgt ctgaaagaga ggataacgtt 120 tacaaggcta aattagccga acaggctgag agatacgatg agatggtaga ggctatgaag 180 aaggtcgcca aattggacct tgagttgacg gtggaggaac gtaatctttt gtctgtcgca 240 tataagaatg taattggagc acggcgagct tcttggagaa tcatctcatc gatcgagcag 300 aaagaggagt ctaaaggagc cgatgacaaa ttggaaatga ttaggcaata tcgacaacag 360 gtcgaaaaag aattaagaga tatttgctca gatattctta acgtgttgga taaacatttg 420 atccccgcag cttcaactgg agaaagtaaa gtgttttatt acaaaatgaa gggagattat 480 catagatact tagcagaatt tgccacaggc acagaccgca aagatgccgc agaacattca 540 ttagtagctt acaaaagtgc aagtgacatc gcaaccacag aattacctcc aacacatccg 600 atcaggctgg gtttagctct aaacttcagc gtcttctact acgaaattct caacagtcct 660 gatagggcgt gcaggcttgc caaggcggcc tttgatgatg ccatcgccga gttggatact 720 ctcagtgaag agagttacaa ggacagcaca cttattatgc agctgctcag ggataaccta 780 acattgtgga ctagtgatat gca 803 // ID GT422743; SV 1; linear; mRNA; EST; INV; 813 BP. XX AC GT422743; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A04 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A04 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-813 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 7359e3acf6e3ac609f980e0ced4ae78a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 4. XX FH Key Location/Qualifiers FH FT source 1..813 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A04" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 813 BP; 256 A; 144 C; 207 G; 206 T; 0 other; gggggtcgtg tctaacctaa aaagttgaga aaaaaacgca ttttaatcaa tccctacagt 60 caaagccgta atgtttggtt tgaaactatg gatttaagtg agcaaattaa cttggtgaac 120 ttgaacagtg aagccccttt aagtgttgaa aatttcgctg ctgtgatcgc tggccaccat 180 acagaggtga ttattagcta ctttagcaac aacacctgct actttatcac gcagtttggg 240 aagatcggca atctctacgc tgtagagagc gaagaagtga aaaacgggat actggctaat 300 gagagggtgt ttaatttgcg cactttgctg ggtggagaca acacagagat agaggcaggt 360 gttcgtttta ttgcggagca actggacgtt aaacagcccc taactgttag tctgacgctg 420 aaaagctaca ggccggagca tttgaagcta atcgtaaagt gtattaagga gtttaaggcc 480 aactagagca gaatggccct agtgaaaagg actaaactag cggagagtta ttttggtcag 540 gttgtgctcg gaccgccagg cagtggaaag actacgtact gtggcaaagt gtatgagttt 600 tacaagaaca agttaaacag acaagtgcag gtggttaatc tagacccagc caatgaaaac 660 atgggctatg ccccgacgat cgatcttatg aatctgatta ctgtagagaa agtgatgaaa 720 aaatataatc taggaccaaa cggcgctttg atgtactgca tggagtactt ggagcagaac 780 ttcgaatggt tattaaagca actagttcaa gtc 813 // ID GT422744; SV 1; linear; mRNA; EST; INV; 796 BP. XX AC GT422744; XX DT 02-JAN-2012 (Rel. 111, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A05 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A05 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-796 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4f1c9a16ced1cda53d9b09ba52d4bb08. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 5. XX FH Key Location/Qualifiers FH FT source 1..796 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A05" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 796 BP; 237 A; 158 C; 181 G; 220 T; 0 other; gggggccgga aaaattagtc aaaaatctat gagccatcgc cattttgcac tgttgtgttt 60 tgtctaccaa gtttattaaa gagtgtccgt ttgcggttaa atttacacta ttttcaacgg 120 gaattagcct tagaataggg ttttgaattc cactaataca tttctgtgca aaaggagttc 180 gctacgatgg ccgtagatga caaggcgtct tctaaggagt tggaccagtg gattgaacaa 240 ctaaacgact gcaagcaatt aacagaaaat caagtaaaaa cgctatgtga aaaggctaaa 300 gagatactct caaaggaatc taacgtgcaa gaagtaaaat gtcccgtaac agtctgcggc 360 gatgtacacg gccagtttca cgacctgatg gaactcttca ggataggtgg acgcagtcct 420 gatacaaatt acctatttat gggcgactac gttgaccgcg gttattactc ggttgagacc 480 gtaacgcttc ttgtcgccct taaagtccgc tataaggaaa gaattacgat tctgcgagga 540 aaccacgaat cgagacaaat cacacaagtt tatggattct atgatgaatg tttgagaaaa 600 tatggcaacg ccaacgtatg gaagttcttt actgatttat ttgactacct accattgacc 660 gctcttgttg acagtcaaat attctgcctc catggcggtt taagcccaag tattgataca 720 ctagatcaca ttcgggcatt ggatcgtttg caagaggttc ctcatgaagg gcctatgtgc 780 gatctgctat ggtcag 796 // ID GT422745; SV 1; linear; mRNA; EST; INV; 781 BP. XX AC GT422745; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A06 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A06 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-781 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; cce28ee3fdd565f731cc70675faa53f4. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 6. XX FH Key Location/Qualifiers FH FT source 1..781 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A06" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 781 BP; 249 A; 174 C; 218 G; 140 T; 0 other; ggggggcccg agtggacata acctcaaata ggtggtgtaa tttggggggc aatgctgcga 60 gccatcagca acttcaaccc cctgcggggg tccagtgcca gtttggctca ttatcactac 120 accggttacg atccgtctct gttgccgccc aagaaggctc atcgcaaacc gcgctgggtg 180 ccggtggcga aaagcaagcg ctttcgggtg ccccaaaggc ctaaaatcga tcaggatgag 240 tatgtggagc tgttgcggct gcacaacaac taccgtaaca acgtgaagtc gctgaggcac 300 tacatgatca aaaagtactg cgacaagttc caaaccgtga ccaatccaga ggagattttg 360 gaggtcttta aggtggacct ggccgaatgt catgcaatca acgaccagtg gaatgctaaa 420 gtcaaagtgg agcgtgagag cagaatagcg agggaactgc aggaagccaa ggagatcgcc 480 agacagaagc ttgaagctga acaggagaaa cagaagcaaa tcctggaact ggctgaggaa 540 attgtcaaaa aggaaaagga ggcctcagct accttcatta cggcagaaaa catcgacgaa 600 gccatcgata atgctctcaa cgacacagtg gactacaact ttgccattga tagcaatggg 660 gagaagctga ggggcagaga gacgcggcct agcgggccgc aggagaaaat cagcgtgcga 720 cgataagtct gtacagtagg ttaatattaa ataaactata tcacaaaaaa aaaaaaaaaa 780 a 781 // ID GT422746; SV 1; linear; mRNA; EST; INV; 761 BP. XX AC GT422746; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A07 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A07 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-761 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 8830ad8bd6fab7f3da4e6f388064745b. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 7. XX FH Key Location/Qualifiers FH FT source 1..761 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A07" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 761 BP; 237 A; 163 C; 199 G; 162 T; 0 other; ggggaggatc aaggcctggg ccgatattca aaaggtctac aagagtcaac gtttccgtgc 60 tggcaaggga aaaatgcgca acaggaggag aattcagcgt aagggacctc tggtcgttta 120 ttatcaagat caaggtctac gcagagcttt ccgcaatatc ccgggtattg atttaatcag 180 tatcgagaag ttgaatctac tgaaattggc gcctggcggt catgttgggc gtttcgtcat 240 ttggacggaa tccgctttcc aaaagctgga caaagtgttc ggaagttgga aagtggcttc 300 cagccacaaa aagggctaca atctgcccga aactaaaatg tccaacaccg atctgtctcg 360 cctgttgaag gctgatgaaa tcaaggcggt cctgcgtcgt ccacagaaga agattgtgcg 420 tcgtgtgcga cgcctcaatc cattgacgaa cgccaaggct atgttgcggc tcaacccata 480 ctctgcggtt ttgaagcgcg aagcaatcct atcaggccag aaaagaagct tggctagaga 540 ggaacttctt gccaagaagc gcgggatcac tttaccggaa acaagtccgg taatcaggtc 600 ggccaagctg caggctaaaa ggagggtcca aatattgaag gcccagaagg caaaggctgc 660 caagccgaaa aaagttgccg cgaaggcacc agctaagaaa taaatatgat gtttcttagt 720 tacgaaataa ttatcaagta agataaaaaa aaaaaaaaaa a 761 // ID GT422747; SV 1; linear; mRNA; EST; INV; 776 BP. XX AC GT422747; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A08 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A08 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-776 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 7713169ccfad67bc49fedceed0d12d2a. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 8. XX FH Key Location/Qualifiers FH FT source 1..776 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A08" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 776 BP; 276 A; 141 C; 161 G; 198 T; 0 other; gggggcagtt ctctataacc agttgcaaaa ttaaacttga agaaaatgac tacatgtatc 60 caaagtgaaa ttaaccagtg ggttgccgat accttcaaag agaatcaatt caaagacctc 120 agtgctgaaa tagttggaac ttcagaacag ggcgaagggt acctaggaaa tataactttt 180 gccaaagtca caggtgtacc attttctgga aaagccaaag aatttcacgt agttattaaa 240 tcgggaaaac gaggggatgg caccacaaat ctatcttcag tccaactggc ctatgaaagg 300 gaaaacttct tctacgataa agcagttccc gctttccaag aaattgagaa aaaatatgga 360 atcaatataa ctgagcaatt attcccgaag tgttttagta ccctttccta cgaagatcag 420 caattgatcg tattgcaaaa cctgaagtct ttagattttg agctttacga cagacaccag 480 cccttcgata tggagcattt gaaagtcact ctttcaaact atgggaaatt tcacgctttg 540 agtttagcat atcgggagca caaacctaag gaatttaagg agctatttag tggactgcca 600 tgtgctatga aaaatattat cgaactgacg aaaaattcag tagatgcagc tgaaaagtgt 660 ctgtacatga tgctagagga aaatcaaaag tttaaggcag ctgaaaaact gaaagagctg 720 cttccggatg gagtcaccgc aaaaattcaa caaatgttaa gaattcaaga gcatga 776 // ID GT422748; SV 1; linear; mRNA; EST; INV; 847 BP. XX AC GT422748; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A09 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A09 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-847 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 1fe6faeb10a27f6b41349d521a44d4e5. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 9. XX FH Key Location/Qualifiers FH FT source 1..847 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A09" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 847 BP; 263 A; 147 C; 171 G; 266 T; 0 other; ggggagttaa aatcggttga atcagaagta ataatcatat aattgtggta cggttgcatg 60 taaatataat ataatttagg ccgcacaatt atttaattta ttattagctg tgaagtggtt 120 gatgacctat atttgaaact ttgaaggaaa aaatctgata ataatgctat aatataataa 180 attacacatc ctattccgaa tttaagtcac aataatcaca atggctaccc gattcgccaa 240 aggcgaagat ccaacggaga ttttacaatt taacttcttg gattcggaag gacgtatcac 300 caaggaatcc attaaatcgg ctttaaaacc cccatatcct gcaaagtatt atataaatgt 360 tcttttgatg gttatagctt tactcgtatt tcttctttgg aactatggta tcatacaggg 420 aacgaaactg ggattcttct gcaatgatcc tttattttcc cacaaatacc ggggggatac 480 agtaatgccc tggatgttag gtgtggcact gggcattatg ctaccaggct tgtatcttac 540 agtagaatgg attaggcaaa gcaatttcga ggctgtattc agcatagagc acatttttga 600 attctatgta tatttcaagc attatatgat tgggcttgta attgttgcag gagtcacaga 660 attagcaaaa accatagtag gggagcacag gccccatttt ttcgacactt gcaggccgga 720 tacgaatctc aattgcacca atggaacgta catcttcgac tacacctgta ctaataaaaa 780 tgttgcaatg ctggattggg ttgatgcttc cagatcgttt cttagtggac attcatcgat 840 cagttgg 847 // ID GT422749; SV 1; linear; mRNA; EST; INV; 816 BP. XX AC GT422749; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A10 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A10 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-816 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5ab9119c1af638e65c6fd9398ae06ec5. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 10. XX FH Key Location/Qualifiers FH FT source 1..816 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A10" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 816 BP; 244 A; 181 C; 203 G; 188 T; 0 other; ggggggcttg ctgaacttgc acaaacggat cttccatttg ttatttggtg atcgatatgt 60 ctcaccgaaa gttcagtgcc cctcgtcacg gttccatggg cttctacccc aaaaagaggt 120 ccaagaggca tcgtggtaag gtcaaggcgt tccctaagga tgacccatca aaacccgtgc 180 atttaaccgc cttcctcggt tacaaagctg gtatgaccca cgtggttcgt gaagctgatc 240 gtccaggttc aaagatcaat aaaaaagaaa ttgtggaagc tgtaaccatt cttgaaacac 300 cgcccatgat tgtagtgggc gtcatcggtt acattgaaac tcctcatggt cttcgcagtc 360 tctgcaccgt ttgggcagaa cacttggccg aggagtgccg taggcgcttc tacaagaact 420 ggtacaagag taagaggaaa gctttcagca gcaatgccac gaaatggaat gatgcgcttg 480 gtaaaaaggc cattgaaaag cagttcaaca aaattatcaa gtactgctca gctgtgagag 540 taatcgccca cactcagatg aaactgctga agcaaaggca gaagaaggct cacattatgg 600 aaatccaaat taatggcggt accattgctg aaaaggtcgc atgggctaga gaacatctgg 660 aaaagcccgt ccctgtcaaa caagtatttg gccaagacga ggttattgat gttatcggag 720 ttaccaaggg taaaggattc aaaggtgtca cctcaagatg gcatacgaag aaattgcccc 780 gtaagactca caaaggtcta aggaaagttg cctgta 816 // ID GT422750; SV 1; linear; mRNA; EST; INV; 858 BP. XX AC GT422750; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A11 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A11 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-858 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 4c97a849910afc8ec860a071aecbe4db. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 11. XX FH Key Location/Qualifiers FH FT source 1..858 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A11" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 858 BP; 220 A; 213 C; 216 G; 209 T; 0 other; gggggatctg tgatttgttt ttgattgata atgtccaaaa tttgttgcta attaggaaaa 60 agtgcattta ttccctactt aatttagcag cttacaatgg atgtggacaa tggagatgga 120 ggaggcgtga gcggctccat ggacatagaa cagtctctgt tgcaacagtt cagttgttta 180 ggaacaactg acaaggatga acttgtacaa cagcttcaga agcttttggg cagtccaacg 240 catctcaatt actcaacggc tgcatttttc ttggacatga acaactggaa cttgcaggcg 300 gcgatttgct cctatctcga cgtggaggcc cccaacacac tgccattgat ggccttagtg 360 tccgatccag atgcctccga gacggaaaac attgagccaa acacccagtt ccaaaaggcc 420 tggcatatca gcaattcggg gacggagagc tggccagtcg gctgctacat ccagtgttcc 480 gatggtgata cttttggggc cgaacgagta tacctgccag ccctacagcc tggccagagc 540 acttatgtga ttgttgcgat gcagagtccc cccatgcctg gaatctacca gagtaaatgg 600 agggcgtgca cgccggcagg ttcttatttt ggagatccga tgtgggtgat tctcactgtg 660 gtagaacagg gcaccattga gctaactgag cagctgtccc actttagcga actgggcgcc 720 atccccccac tggtacccca agtgccgaat cccttcattc cccactacat acatagagaa 780 aacaaggatt ccgccccacc cgatggacaa ttctagtact gcagggctcg gcaaaagtgc 840 tccagattgt taactgaa 858 // ID GT422751; SV 1; linear; mRNA; EST; INV; 802 BP. XX AC GT422751; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A12 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A12 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-802 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e555d1a3a21774326718c9ee70e6a7b6. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 12. XX FH Key Location/Qualifiers FH FT source 1..802 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A12" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 802 BP; 260 A; 170 C; 180 G; 192 T; 0 other; gggggagaaa aacatgcgaa agtttaggaa tttccaaaca aacgccccga caaattaagg 60 aaattgactg aaatggttga cgctccgcaa tacccaacta tcaaccgaga ggcaccgaat 120 atactagtaa cgggaactcc cggtgttgga aagtccactc tatgtaagaa attggccgaa 180 gtgactgcat tgcgttggtt ggaaatctct aaaattgcca aggataataa ctgtttgcag 240 gagtacgacc ccgaattcga tacacatgtg ctggatgagg ataagctaat ggacggcctg 300 gaggaagtta tgagcatagg gggaaacata gtggactatc acagcagcga atttttccca 360 gagcgatggt tcgatgtagt gttcgttctc acggccgaca ctagcagtct gtatgatcga 420 ctcaatttgc gtggctattc aggcaaaaaa ctagaaagca acatcgaatg tgaaatcttc 480 aaaatatgtc tagacgaagc aaagcaattt tacagaaccc agatagtaca ttcactggag 540 agtataaatg aagcacagct agaggaaaac gttcagaaaa tatgccaatg gatacaaatg 600 tggctcaaca atcgagaaat gaatccccaa ttaaacagtt aggctttatt agacttaaga 660 ctcaccttag cctattcata atgtcaactc ccgcctggtg ccgaacgcaa ctggtcacat 720 aatccaaagg aatctgcttt ttctgtttgc agtccctttt gatatcctgc actgtgtgct 780 cgtccctcaa cacatacggg ca 802 // ID GT422752; SV 1; linear; mRNA; EST; INV; 781 BP. XX AC GT422752; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A13 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A13 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-781 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 20d0622653e87e320ea4e26b57b0585f. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 13. XX FH Key Location/Qualifiers FH FT source 1..781 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A13" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 781 BP; 236 A; 191 C; 184 G; 170 T; 0 other; ggggagcgtc ccaaaaacaa gatagatttg ggattaccgc tggagtgact actaagaaag 60 tgagtgaaaa agtgcccaaa ctaccgactt tcttgtccga actgaacgaa gcatacgtgg 120 aacataaacg agagccgacc atggcgtcgg aacagttttc actatgttgg gacaattttc 180 acaagaacat gagttccggc atgaacgcgc tgctggaaca cgaagaccta gtggatgtga 240 ctttggctgt tgagggccgt ctattgaagg cccataagat ggtcctgtcc gtgtgcagcc 300 cctatttcaa agaactgttc aaatcaaacc cctgtcagca ccccatagtg tttatgaaag 360 acgttagcta tgtggcgatg agtgatttgt tacaattcat gtaccagggt gaggtacaag 420 tgagccagga caatttaacc actttcatta agactgccga ggccttacaa atcaaaggac 480 tcaccgggga cggaaacggt tcaactgatg ccgagaccga atctctgcct gaaaaacctg 540 cgacgcgtca tactgaggag tcgtacaagc caatgtccag accaaaaaaa cagccactca 600 ctccttcagt gactacaagt cccaatcaaa gtgcaaagcg ggcgcgactt acctcgaatg 660 attcgcagtc atcgcttacc cctattacga aaacggaacc cgcttcggca caaactgcaa 720 ataccgattc agcggttcaa tttaaagtgg aaccctacga ccagaatcag tcagtgttga 780 c 781 // ID GT422753; SV 1; linear; mRNA; EST; INV; 813 BP. XX AC GT422753; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A14 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A14 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-813 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; d7036712788eb487ad63f2b8ddfa0961. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 14. XX FH Key Location/Qualifiers FH FT source 1..813 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A14" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 813 BP; 244 A; 192 C; 232 G; 145 T; 0 other; gggggaggac agcgtacgtt tcccaagcgc cgtcaggaat aaaggtaccc cttgtttctt 60 attcttgcta gaccggacac caaccagcca ccaccatgga cgcgatcaag aagaagatgc 120 aagcgatgaa gcttgagaag gacaatgcgg ccgataaagc cgatgccatg gaagggcagg 180 cgaaggatgc caattcccgc gtggacaaac tcaatgaaga actgcgcgat cttcagaaga 240 aactgtcgca ggtcgagcag gacttggtca gcaccaagaa ccacctggac caagccaaca 300 aggatttgga agaaaaggac aaggctctgg tcgctgctga atctgacgtc gctgcccaga 360 acaggaagct gcagctgatt gaggaggatc ttgagcgctc tgaagaacgt ctcgccactg 420 ccaccaccaa actggctgaa gcttcgcagg ccgctgatga aagctcacgt atgtgcaagg 480 tgttggagaa ccgctcccaa caagatgagg agcgcatgga ccagctgacc aaccaattga 540 aagaggcccg tctcttggct gaggatgccg acaacaaatc ggacgaagtt tcccgtaagc 600 tggccttcgt tgaagacgag cttgaagtag ctgaagatcg tgttaaaggt ggtgatgcca 660 agatcatgga actggaagaa gaactgaagg tcgtcggcaa ttctctgaaa tctttggaag 720 tgtccgagga aaaggccaat caaagagtgg aagaattcaa gaaacaactg aaatccttga 780 ctgtcaagct aaaggaagcc gaagctcgtg cgg 813 // ID GT422754; SV 1; linear; mRNA; EST; INV; 832 BP. XX AC GT422754; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A15 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A15 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-832 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0427854040236dedf4ec56892c579eac. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 15. XX FH Key Location/Qualifiers FH FT source 1..832 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A15" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 832 BP; 259 A; 181 C; 202 G; 190 T; 0 other; ggggctcttg cagttcttag tccaacatgg ccgtaggaaa aaataagggt ctctccaaag 60 gaggtaaaaa gggagttaaa aagaaggtag ttgacccctt cacacgcaaa gaatggtatg 120 atgttaaggc cccgtctatg ttcaccagcc ggcaagtggg taaaaccctc gtcaaccgta 180 cccaaggaac caaaatcgct tccgaaggtt taaagggtcg tgtctttgaa gtcagtttag 240 cagatttgca aaacgacaat gatgctgaac ggtctttcag aaaattcaga ctcattgccg 300 aagatgtaca aggtcgttat gtattaacaa acttccatgg aatggatttg accaccgaca 360 agctgaggtc gatggtgaaa aaatggcagt ccctcatcga agcttcagct gatgtgaaaa 420 cctcagatgg atatttgttg agagttttct gcattgggtt cacccaaaaa gatacacagt 480 tttcgcagag gaaaacttgc tacgctcagc atacacaaat caggcaaatc cgcaaaaaga 540 tggttgagat aatcacccgt gacatccaag gctcggacct caaggaagtt gtaaacaaat 600 tgttgcccga cagtatcgcc aaggatattg aaaaggcctg ccagggaatt taccctcttc 660 acgatgtctt catccgtaaa gtgaaggtgc tcaaaaagcc ccgatatgag ctttcgaagc 720 tattggaact tcacggagac gccaaaggca ctacagaagc atcaggggat gctggagcgc 780 aggtggcgag acctgagggt tatgaacctc cagttcaaga atccgtctaa tc 832 // ID GT422755; SV 1; linear; mRNA; EST; INV; 587 BP. XX AC GT422755; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A16 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A16 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-587 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 74a839611d749c1c2a54f991f6b02838. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 16. XX FH Key Location/Qualifiers FH FT source 1..587 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A16" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 587 BP; 191 A; 141 C; 135 G; 120 T; 0 other; gggggagcca attgtcactt gtcattggaa ccatcttttc ctgttcgtgc acgccgcgta 60 aagaccttca acatgtcctc acacttggtt tggagcatca tccgcaacaa caacgccttc 120 ttggtggaga agcgcaatat ttccaagcct ttcagtactg agcccaacaa cttaaccaac 180 ctgaactcgt acaggtacaa tgggctggtc cacaagaaga cgttggccat tgctgatggt 240 cccgacagaa agggcttcac gctgacgtac aaaaaggcca agaagcagaa taagcccagg 300 cagaatttgg tgaaaaggac gttcaagtcg aagcccaggc gctcgttgta caagctgaag 360 aacttcatgc aggccaacaa gtacaggacg gacttgacca aggctgccct taggagggcc 420 agcgcagtcc tgcggtcgca acggccccag cctgccaaga agcaaaagcc taaaaagcca 480 gaataaataa ttttgtacga gtaagcacag caaaaatccc ttaaattttt gaactggtct 540 tgctgaaatt aataaataat ggtatggata aaaaaaaaaa aaaaaaa 587 // ID GT422756; SV 1; linear; mRNA; EST; INV; 541 BP. XX AC GT422756; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A17 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A17 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-541 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 6944aeb1008b44110f8c4687290d9423. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 17. XX FH Key Location/Qualifiers FH FT source 1..541 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A17" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 541 BP; 169 A; 114 C; 148 G; 110 T; 0 other; gggggacact tcggcgagaa acctcgacaa aagattttcc tcaaaaatgt ctgacgttga 60 aacagatgat gctcccgcgc caacccccgc acccgcaccc gtcagtggtg gccccatgga 120 cgtgaaccag gccctgcagg aggtactgaa gaaggcgttg attcacgatg gtgtcgttca 180 tggactgcac gaggctgcta aggccctgga caagaggcag gcggttttat gtgtgttggc 240 tgagaactgc gatgaaccca gatacaagca gttggtgcag gccttgtgtg cggagcacca 300 gatcgacctg attaaggtgg acaacaacaa aaaactgggc gaatggtcag gactatgcaa 360 aatcgacagc acggggaagg cgaggaaagt cgttggcagt tcgtgtgtag tcatcaagga 420 ctttggataa gagtctccag cccttgacgt ccttaaggaa tacttgaaaa acaacaaata 480 attataaggt tttttatgta atataaggac aattgaaaaa tgcgaaaaaa aaaaaaaaaa 540 a 541 // ID GT422757; SV 1; linear; mRNA; EST; INV; 842 BP. XX AC GT422757; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A18 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A18 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-842 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a0e5f97a11c0662e54a0cc54c072642b. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 18. XX FH Key Location/Qualifiers FH FT source 1..842 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A18" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 842 BP; 264 A; 150 C; 199 G; 229 T; 0 other; ggggaacaga gcttgtgctt gcgtactttc aacaatgaag tgttttattt taattgggct 60 aataggcctt ggttgggtag tagccattct aggggcctct tgtacggtgt caaactatta 120 cgatgtttca acggcagctg aaacctgcga cacattgatt ataggagatt tggtaattcc 180 tgcagaaacc actctggaaa ttaatttgaa agatggtgca aagttgattt ttgacggtca 240 tttaacacat gaagtggctg agtggactgg cgacttagtt cacatcaagg gaaagaatgt 300 tgaaatatct ggaacagcaa accatctgct tgacggtctg ggtcctcaac attggaaagg 360 agcaggcgaa tcaacaatat tgcggcccaa gtttatgcga attcaggtag aagattccac 420 agtgaaggac ttgaacatca agaactgtcc caacaattgc gtcgttttgt ccaagtctac 480 taacttgctt gtaacgaaca ttaacattga taactatgat ggatacccag gagttatcgg 540 agcaaactat gcgaaaaaca ccgatggatt tgatgtggct ggaggcagcg gaatcagagt 600 ggaaaactca gtagttatca accaggatga ttgcgttgca gttaacggag gatcgaatat 660 ggttttcgaa aatttacact gcaatggctc ccatggattg agtttctcga tcaaaaatgg 720 ggacgtgtcc gatattcaat tcttaaactc caaagttgag tattccaaca atgtaataca 780 catcaaaact cacaacgatg ggggatacgg gatcaataaa aaattgtgct ctataaaaat 840 at 842 // ID GT422758; SV 1; linear; mRNA; EST; INV; 761 BP. XX AC GT422758; XX DT 02-JAN-2012 (Rel. 111, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A19 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A19 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-761 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; cf1303bc85bb41e71075a6183d0bbf2d. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 19. XX FH Key Location/Qualifiers FH FT source 1..761 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A19" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 761 BP; 221 A; 156 C; 171 G; 213 T; 0 other; gggggacaaa tccttgtcac tgaaagtgaa aaatggatac cccaggcaga cctaaattgg 60 ccgaaactcc tctggaactg ctcaactctc ctggcattga acctggagta ctcaatgtgc 120 tctacctgcc cattgtgggg ggagttttag gattcgtttc gattcttgta gcgaattggg 180 ctggaaaaag accagtgatg agtggcatcc acaagcacat tttgggcctg agtgcaggtg 240 tgggattcgg ggttgtaacg gataaataca gaaacgaaca gctggcgaca agagacgcaa 300 tatttcgcca ttacatcgag ctgcaccccg aagacttccc cccttatgag cgtgtaaaaa 360 tcagggatgt tttcggcccg tggagaccaa ttcgttgatt tgttttgtat gtaagaaata 420 aactagttct agtaggtttg ctttgctttg agtttccaat ggaaaatccg aagaaaaaac 480 ctcctctagt tcatgcttta attatttatt tgtggcttaa aggagtttgg attaaacttc 540 taaaggtagt atacacatgt aaacagcttg aaataacagt atcaagtttg tacattcagt 600 atttgttaaa aggacaacat gggaaactga acaagtagaa taaccattgg aaaatcgctc 660 catgttgcca atctccggcc tttgcaactt gactgtgcga aatttgcgct cgatttctca 720 tgcaaattgc gcccagcatc acttttcccc aataaaaacc t 761 // ID GT422759; SV 1; linear; mRNA; EST; INV; 795 BP. XX AC GT422759; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A20 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A20 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-795 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; ca3c7b96a05fe1a3e830700efd9a92df. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 20. XX FH Key Location/Qualifiers FH FT source 1..795 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A20" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 795 BP; 254 A; 172 C; 163 G; 206 T; 0 other; gggggaacac cacatttgcc ttgcttaggt attctaacct tcttttaaca gttggaaaca 60 ctatggatat ttttaggaaa ttgagtcgtt actttccata tttaacatcg agatgaagta 120 tgaacagaaa ccccaaaatc aggtaacttt ctccatataa accgtagcca ttagctgaaa 180 cttgtaggta tgcaggagcc cttttggttg gttaagtttt catcagaatc ggacgccaaa 240 ctactaacca gcaggtggca agtggcctaa ggcagctctt aaagaagctc tcattgaaaa 300 ccaggaagtt tattggaaac accagcatgg atccgcagct ctcattgcta atggcaaatc 360 aggctcaagt gaatagtggc agtattattt tggacccatt tgtaggctcc ggatctttgt 420 tggtggctat gttttcggat ctgatatcga ctatttgatg ttacatggga agacaaaacc 480 gaccaggatt aaacagcaga caatagccaa ggacgagagt atcaaagcca acatgaaaca 540 atacaactta ggccacaaat acattgatgt gttaatcaac gacttcagcc tgcccttttg 600 gagagattct atggaattcg atgccataat aacagaccct ccttatggaa tcagagaagc 660 caccgaaaga gtgggaacgg agaagcaaaa ttacatagtc agcgatgaac atttgcccac 720 tcacgtaccc gcaaaaatcg aatacaaaat tcccagcatc ttccgacact tgctgatatt 780 cccatccaac acttg 795 // ID GT422760; SV 1; linear; mRNA; EST; INV; 857 BP. XX AC GT422760; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A21 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A21 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-857 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e32fe5cf79623be4bce8f2517f94c270. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 21. XX FH Key Location/Qualifiers FH FT source 1..857 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A21" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 857 BP; 248 A; 150 C; 166 G; 293 T; 0 other; ggggaatcac aaaagtactt gtcgctgtag aggcagttta ggttcgattt atttcagaaa 60 aatacatttt ttaaaacgtt gagttcagtt tacataagat aagaaattaa gaattttcca 120 aaatggtaga tattcatgcc gatattaaaa atcgactatt tacaataacc aggatggggg 180 gcaacgagtt gcttacaccc ggcacggaac tcaccgagct aaaatttgag gggttcttag 240 attggttcgt ttgtattact gatccaaaag tgcattggaa atatgaaccc acttatatta 300 tttcccagac agttgttgta cttggagggc tgttcaccct gatacatgca atagttagag 360 gaggtcgtct tccatggtta tggttcggaa taactttgca tggaatattt gtggaaattc 420 tatgctacat tattccagat gtagacaatt tctggcattc tcaaactcca ataatattct 480 tcggcagacg acttcccttg catataattc tagtttgcgc gtattttgcc agagcaaagg 540 aaaatggttc aaaggaaaaa gctttttaaa agaattcttc gcaactttct tggcgtcttt 600 attggggccc attggtggct tgcctctctt cattccgata taccatccat tgcatgatgt 660 ttttaatatt cacagtgaag ttacctattt cattcttttg actatcttct tactcctaat 720 ttggagtggc gataggatga gtagatcgaa cgaagttaat tggaaaattc ccaataaaac 780 acattggaca acgttttttg ttatattgca tctggtactt cattatacta ccttctggat 840 tattccgatt ttattta 857 // ID GT422761; SV 1; linear; mRNA; EST; INV; 862 BP. XX AC GT422761; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A22 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A22 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-862 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; a82a5153a35e1d43dae1df6d486a159b. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 22. XX FH Key Location/Qualifiers FH FT source 1..862 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A22" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 862 BP; 335 A; 159 C; 191 G; 177 T; 0 other; ggggatcaaa gtcaatgtaa tctagacgca aaccaagcca ttgaaaacat taaaaagcct 60 actaaaaatc aagcttcgta cataccgaag aaatgaaaca agctgcggcc cagctgcgat 120 tgtcatgccc ataacatcct tgtttgttga attttggtag aatccaccgt cttcagaacg 180 aaaaataatg tcgtttttcg gatccccctt atcaaactcg tacaataatc acgagtatga 240 ctttgaagaa gacaaactag atgaggatga ggagaaagaa cacgatcgta tggatggcag 300 tgatgaagac actgaagatg ccagtgaaac agatacagga cgtcaagagg aacaattgga 360 aatcaaagag caaatgtacc aggacaaact agcgactctt aagaaaaaat tgacccagct 420 aaaagatggc tcgcacggcg attacaacaa aaaagtcaag agattggaat cacagtataa 480 cgaaagattg aagttgaatg atatctataa agattatatg tatgagtaca taaagaggga 540 ttatgctcaa gagatcaaag cggcagccaa ggacttcgag gaaaagaaaa tcgatttgag 600 ggaaacattg atctctgact tggaagataa gaaacgtaac atagaaacgg aaagatcttc 660 tatggagctg actggagact ccatggaggt aaaacctgcc atgactagga aactgagaag 720 gcgcccaaat gagccaattc ccatgcccga aaaaaggcga aaagccccaa cagcccaaat 780 aacctacttg ctggatgata aagaagctga caacgatttg agaatgatta aagaaaaagc 840 tgttctgccc gttaagaaat tg 862 // ID GT422762; SV 1; linear; mRNA; EST; INV; 811 BP. XX AC GT422762; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_A24 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_A24 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-811 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 0df675ae9a5e9211e94f9df802f990b4. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: A column: 24. XX FH Key Location/Qualifiers FH FT source 1..811 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_A24" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 811 BP; 206 A; 212 C; 210 G; 183 T; 0 other; gggatcgatc aggacgagtt cgggattatt ccttcgaaac tgatcgacgc cttggagaac 60 tgtaaagcct acacggagga aggcggaggg aaacgaatgc caaaggccat ctacctcaat 120 ccgaccggtt cgaacccgac aggcattaca atgtccttag agcggaggca ggaaatctat 180 gcgatttgct gcaaatacaa catcctgatc ctggaagatg acgcctactg cttcctacac 240 ttccaagagg aggagcccgt ctccttcctg tccctggacg tggaagggcg agtgatcagg 300 ttcgactcca tgtcgaaggt tttgtcctct gggctgcgcc tcgcttggtt aaccggcccc 360 aaacaattgg tcagcaacat tgagctgcat attcagagcg acatgctgca ttccagcact 420 ttgtctcagg ttatcttcga caatctgatc tctgcctggg gcttcaatgg tctcctttcg 480 cacttcagct acgtgagacg atactatcaa gcccgacgtg acttcaccat agcggccatg 540 gacaagcatc tgaagaacat ctgcgaatgg acagtgccca caggtggaat gttcgtctgg 600 atcaaagtgc atggagttgc cgatgtctac gacatgctaa tgacccgagg gctgaagaag 660 aacatcactt ttgtgcctgg acacgctttt atggccgatc ccactcaggc gtgccagtac 720 atccgggcat ccttcagcaa agcgagcttg aagcaaatcg acaaagccat gaagctactg 780 ggcgagttga taaaggaaga acacttgctg c 811 // ID GT422763; SV 1; linear; mRNA; EST; INV; 776 BP. XX AC GT422763; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_B01 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_B01 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-776 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; e6ddbd460c086c2f6df6b4db2b009b93. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: B column: 1. XX FH Key Location/Qualifiers FH FT source 1..776 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_B01" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 776 BP; 219 A; 156 C; 180 G; 221 T; 0 other; ggggacagct ggaaaacata accataacct acccatgttt tgttatttta attttcagtt 60 ttttaacagt gttttatttt ttaaattgat atgaacgagc tggaatttaa gtgtggaaac 120 cctggagcat caaagtttgg caatttcatc aattactatc agttccaccc tccagatgag 180 cggatttcca tgttggcccc tgaaatatgg agttctgcct tcaaagcgga caggggctgc 240 gctttagacg ttggatgcaa ctcaggggat ctcacagttg cactcaagga tttcctcacg 300 aacgttgtgg atacccccat ttccatatta ggcgtggacg ttgacccagt tttaattcga 360 agggcgtgtg agaagcaatc cccaagaaac cttacattta agtgtctcga catcatgtca 420 gacaccgggg atattttcac agacttccta aagccccgca atcgcactag gtttgacata 480 gtcttctgtt tttccataac tatgtggatt cacttgaatc atggcgatga tggattgagg 540 aaatttctgg caaaagttgc cagtctttca gactttctgg tcatagaacc tcagccctgg 600 aaatgctaca gaacggctgt caagaggctc aagctgggca aggctgagtt tcccaacttc 660 aaagcactgc aacttagaaa taacgtagaa agtgaaatag agacgtttat tgttgataaa 720 tgtggcttgg agaaggttgg aaagagtgga ttgacaaaat ggggcagaaa gctgct 776 // ID GT422764; SV 1; linear; mRNA; EST; INV; 784 BP. XX AC GT422764; XX DT 18-NOV-2009 (Rel. 102, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_B02 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_B02 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Dendroctonus. XX RN [1] RP 1-784 RX DOI; 10.1016/j.ibmb.2012.03.010. RX PUBMED; 22516182. RA Keeling C.I., Henderson H., Li M., Yuen M., Clark E.L., Fraser J.D., RA Huber D.P.W., Liao N.Y., Docking T.R., Birol I., Chan S.K., Taylor G.A., RA Palmquist D., Jones S.J.M., Bohlmann J.; RT "Transcriptome and full-length cDNA resources for the mountain pine beetle, RT Dendroctonus ponderosae Hopkins, a major insect pest of pine forests"; RL Insect Biochem. Mol. Biol. 42(8):525-536(2012). XX DR MD5; 5d24df7fc90e8ae261915ab748884e99. XX CC Contact: Christopher I. Keeling CC Genome BC-Genome AB The Tria Project-Mountain Pine Beetle System CC Genomics CC University of British Columbia CC Michael Smith Laboratories, 2185 East Mall, Vancouver, BC V6T 1Z4, CC Canada CC Tel: 6048275314 CC Fax: 6048222114 CC Email: ckeeling@alumni.sfu.ca CC Plate: DPO1110 row: B column: 2. XX FH Key Location/Qualifiers FH FT source 1..784 FT /organism="Dendroctonus ponderosae" FT /lab_host="E. coli DH10B" FT /mol_type="mRNA" FT /sex="male and female" FT /clone_lib="LIBEST_025352 Dpo11-EAdult-MGFB-Fed-Norm FT (DPO11)" FT /clone="DPO1110_B02" FT /tissue_type="midgut and adhering fat body" FT /note="Vector: pDNR-LIB; Site_1: Sfi I; Site_2: Sfi I; FT Un-attacked and mountain pine beetle-attacked Lodgepole FT pine (Pinus contorta) trees were felled on June 16, 2008 in FT Jackman Flats Provincial Park, BC, Canada (approx. N 52 FT 56.308' W 119 22.708'). Attacked bolts with waxed sawn ends FT to reduce dehydration were kept in laboratory emergence FT cages at room temperature. The un-attacked bolts with waxed FT sawn ends to reduce dehydration were stored at 4oC until FT several hours before use. Adult beetles that emerged July FT 7-14, 2008 were sexed and placed on moist filter paper at FT 4oC until use. On July 22, females were placed headfirst FT into 4 mm holes drilled into the bark of un-attacked FT Lodgepole pine bolts and held in place with mesh. After 24 FT h of feeding, the mesh was removed, one male was placed FT into the same hole, and the mesh replaced. The bolt was FT placed vertically at room temperature and ambient natural FT light for an additional 40 h of feeding before the beetles FT were manually extracted from the bolt. The anterior midguts FT and adhering fat body from a total of 34 beetles were FT dissected under water and immediately frozen in liquid FT nitrogen. A normalized, full-length enriched, directionally FT cloned library was prepared with the Creator SMART cDNA FT Library Construction Kit (Clontech, Mountain View, CA, USA) FT and the TRIMMER-DIRECT cDNA Normalization Kit (Evrogen, FT Moscow, Russia). Total RNA (106 micrograms) was extracted FT from frozen midgut/fat body tissue of 34 insects (weight FT undetermined) using the Qiagen RNeasy Mini Plant Kit. First FT strand cDNA was prepared from 1 microgram of total RNA, FT SuperScript III reverse transcriptase (Invitrogen), CDS-3M FT primer (Evrogen), and the SMART IV Oligonucleotide FT (Clontech). Second strand cDNA was prepared by LD-PCR with FT Phusion Hot Start DNA Polymerase (Finnzymes, Espoo, FT Finland). This cDNA was then normalized with FT duplex-specific nuclease and amplified following the FT TRIMMER-DIRECT protocol, digested with SfiI, size FT fractionated, and cDNA larger than approx. 500 bp was FT ligated into SfiI-digested pDNR-LIB (Clontech). The FT ligation mixture was transformed into ElectroMAX DH10B T1 FT Phage-Resistant electro-competent cells (Invitrogen). Titre FT of the primary library (1 mL) was approx. 6x10^5 cfu/mL and FT was not amplified." FT /db_xref="taxon:77166" XX SQ Sequence 784 BP; 255 A; 125 C; 145 G; 259 T; 0 other; ggggacctgt tttagttcac attcaaactt ggtttgaagc gaaaattgaa aaattaaaat 60 gtccatctcc actaaatcat tttttgtata tttagccgta gctttagtta ttctatctgt 120 gcttactccg gacgtagagt gcagaagaaa tgttttgcgc ggtaggaaaa ccgtaaccag 180 gcggtatttg cgaccaatgg caattccagc ctgggcagta atcgttttgg taggattggg 240 acaattaatc gtcggagcat tgctttatgt attaatgaag aagctaatag ttgatcctcc 300 cgtgcaagga aactacagta tggcagccac tgacgacgta taatcattgc acatgtttta 360 ttattgtttt cctgtacagg gtgggataac aaatctattt gcgggtataa tagcttaaaa 420 ttattcttac atgttttaag aaactcatcg tatgcctatc tttgtaatag aaatgtaatt 480 tttgccacca ttcagcagag gttaaattta ctgttttgat tcaaaacaac cgtaaatatt 540 tatagtagtt aatttacttg aaacttctgt taaagcaccc ggaatatatg actgcactgg 600 tttggcagaa acgtaaataa taccagaaat tttctttgaa ttgtgatgaa agcatctaat 660 gtgttgtgta tcacgaacgg ttttattaat caccctcctt aatttaaatg atttatattt 720 gtacattata aagatcatag gaaaaacata caaaatacac gtggtcatca taaaaaaaaa 780 aaaa 784 // ID GT422765; SV 1; linear; mRNA; EST; INV; 821 BP. XX AC GT422765; XX DT 02-JAN-2012 (Rel. 111, Created) DT 29-JUN-2012 (Rel. 113, Last updated, Version 3) XX DE DPO1110.C21_B03 Dpo11-EAdult-MGFB-Fed-Norm (DPO11) Dendroctonus ponderosae DE cDNA clone DPO1110_B03 5', mRNA sequence. XX KW EST. XX OS Dendroctonus ponderosae (mountain pine beetle) OC Eukaryota; Metazoa; Ecdysozoa; Arthropoda; Hexapoda; Insecta; Pterygota; OC Neoptera; Holometabola; Coleoptera; Polyphaga; Cucujiformia; Curculionidae; OC Scolytinae; Den