中国农业科学Issue(22)：4808-4815,8.DOI:10.3864/j.issn.0578-1752.2013.22.019
家蚕整合素基因BmintegrinαPS3的鉴定及亚细胞定位
Identification and Subcelluar Localization of BmintegrinαPS3 from Silkworm (Bombyx mori)
摘要
Abstract
[Objective]BmintegrinαPS3 was cloned, analyzed, and overexpressed either in Escherichia coli or silkworm cell line to provide a theoretical basis for further studying the function of BmintegrinαPS3 in silkworm.[Method]The BmintegrinαPS3 cDNA including the complete ORF sequence was cloned by RACE method. The spatial-temporal expression profile of BmintegrinαPS3 was investigated by RT-PCR. Bmintegrin αPS3 protein was overexpressed using prokaryotic expression. In addition, BmintegrinαPS3 was constructed into a transit expression vector, and transfected into silkworm embryonic cell line to analyze the subcellular location of Bmintegrin αPS3.[Result]Bmintegrin αPS3 cDNA consisted of 2 895 bp, and encoded 965 amino acids containing the conserved integrin alpha domains, which was predicted to be a transmembrane protein. BmintegrinαPS3 protein from E. coli was overexpressed and purified using affinity chromatography, and the purity reached more than 99%. In addition, the overexpressed BmintegrinαPS3 protein in embryonic cell line was located in cytoplasm and membrane. RT-PCR results showed that Bmintegrin αPS3 mRNA was specifically and continually expressed in silkworm hemocytes.[Conclusion]The Bmintegrin αPS3 cDNA was cloned and its expression patterns in silkworm were analyzed. The subcelluar localization of recombinant BmintegrinαPS3 protein was investigated in silkworm embryonic cell line. The Bmintegrin αPS3 protein was overexpressed in E. coli and purified for further antibody preparation.关键词
家蚕整合素基因/克隆/时空表达/原核表达/亚细胞定位Key words
BmintegrinαPS3/cloning/spatial-temporal expression/prokaryotic expression/subcellular localization引用本文复制引用
谈娟,张奎,徐曼,陈思源,崔红娟..家蚕整合素基因BmintegrinαPS3的鉴定及亚细胞定位[J].中国农业科学,2013,(22):4808-4815,8.基金项目
国家重点基础研究发展计划“973”项目（2012CB114603）、国家自然科学基金项目（31172268）、中央高校基本业务经费重点项目 （）
