title: CONSTRUCTION OF SILKWORM MIDGUT cDNA LIBRARY FOR SCREEN AND SEQUENCE ANALYSIS OF PERITROPHIC MEMBRANE PROTEIN GENES.
abstract: Silkworm is an important economic insect and the model species for Lepidoptera. The midgut of silkworm is an important physiological barrier, as its peritrophic membrane (PM) can resist pathogen invasion. In this study, a silkworm midgut cDNA library was constructed in order to identify silkworm PM genes. The capacity of the initial library was 6.92 × 10(6) pfu/ml, along with a recombination rate of 92.14% and a postamplification titer of 4.10 × 10(9) pfu/ml. Three silkworm PM protein genes were obtained by immunoscreening, two of which were chitin-binding protein (CBP) genes and one of which was a chitin deacetylase (CDA) gene as revealed by sequence analysis. Three genes were named BmCBP02, BmCBP13, and BmCDA17, and their ORF sizes are 678, 1,029, and 645 bp, respectively; all of them contain sequences of chitin-binding domains. Phylogenetic analysis indicated that BmCBP02 has the highest consensus with Mamestra configurata CBP at 61.0%; BmCBP13 has the highest consensus with Loxostege sticticalis PM CBP at 53.35%; BmCDA17 has the highest consensus with Helicoverpa armigera CDA5a at 70.83%. Tissue transcriptional analysis revealed that all three genes were specifically expressed in the midgut, and during the developmental process of fifth-instar silkworms, the transcription of all the genes showed an upward trend. This study laid a foundation for further studies on the functions of silkworm PM genes.
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Yi-Jun
authors.initials: YJ
authors.lastname: Zhou
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xue
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Yang-Yang
authors.initials: YY
authors.lastname: Li
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Fan-Chi
authors.initials: FC
authors.lastname: Li
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Ni
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Wei-De
authors.initials: WD
authors.lastname: Shen
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Zhi-Ya
authors.initials: ZY
authors.lastname: Gu
authors.affiliation: School of Basic Medicine and Biological Sciences, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
authors.affiliation: National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Wei-De
authors.initials: WD
authors.lastname: Shen
authors.affiliation: National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Zhi-Ya
authors.initials: ZY
authors.lastname: Gu
authors.affiliation: National Engineering Laboratory for Modern Silk, Soochow University, Suzhou, Jiangsu, P.R. China.
authors.forename: Bing
authors.initials: B
authors.lastname: Li
mesh_terms: D000581:Amidohydrolases / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D012831:Bombyx / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics; D041981:Gastrointestinal Tract / Q000378:metabolism; D015723:Gene Library; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D007814:Larva / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D000581:Amidohydrolases; C019866:chitin deacetylase
keywords: Bombyx mori; chitin; peritrophic membrane