烟夜蛾(Helicoverpa assulta Guenée)滞育激素基因在大肠杆菌中的表达  

Expression of diapause hormone gene of orinental tobacco budworm (Helicoverpa assulta Guenée) in Escherichia coli

作　　者：丁矛[1] 杨效文[1] 郭线茹[1] 张蕾[1] 高玉红[1] 马继盛[1] 

机构地区：[1]河南农业大学植物保护学院,河南郑州450002

出　　处：《河南农业大学学报》2003年第3期219-223,共5页Journal of Henan Agricultural University

基　　金：中国烟草总公司资助项目(无编号)

摘　　要：采用RT PCR技术从烟夜蛾(HelicoverpaassultaGuen啨e)滞育蛹中扩增到滞育激素(diapausehormone,DH)基因cDNA,将其与pET 30a(+)质粒连接后转化至大肠杆菌BL21中,测序结果表明,重组质粒插入片段编码序列完整,读框正确;重组工程菌用IPTG诱导表达后,SDS聚丙烯酰胺凝胶电泳检测到一条约21kDa大小的外源蛋白,它与DH的大小相应.DH基因原核表达载体的成功构建为大量获取DH重组蛋白、研究DH的结构及功能奠定了基础.With diapause pupae of oriental tobacco budworm (Helicoverpa assulta Guenée), a doublestrand cDNA fragment of DH (diapause hormone) gene was obtained from RTPCR amplification. The fragment was linked with pET30a(+) plasmid and then the recombinant plasmid was transformed into Escherichia coli BL21. Sequence analysis showed that the insertion fragment of recombinant plasmid was complete and the reading frame was correct. After being induced by IPTG, the recombinant engineering bacterial strain expressed a new protein; its molecular weight was about 21 KDa by checking with SDS polyacrylamide gel electrophoresis. The successful construction of prokaryotic expression vector established foundation for obtaining DH recombinant protein and researching the structure and function of DH. 

关 键 词：烟夜蛾 滞育激素 大肠杆菌 重组蛋白 基因表达 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治] Q786[农业科学—植物保护]