福氏志贺氏菌毒力基因ipaC在大肠杆菌中表达的初步研究  

Expression of Shigella flexneri gene ipaC

作　　者：孙素霞[1] 王红[1] 王勇[1] 向前[1] 吴爱军[1] 俞守义[1] 

机构地区：[1]第一军医大学热卫系分子流行病学实验室,广州510515

出　　处：《中国人兽共患病杂志》2003年第6期29-31,41,共4页Chinese Journal of Zoonoses

基　　金：全军医药卫生"十五"指令性课题 (编号 0 1L0 5 0 )

摘　　要：目的　构建福氏志贺氏菌毒力基因ipaC与质粒pET32a的重组质粒 ,转入大肠杆菌中表达 ,并检测表达产物的活性。方法　PCR扩增福氏志贺氏菌毒力质粒中ipaC基因 ,克隆到质粒 pET32aT7启动子下游 ,转化感受态大肠杆菌BL2 1(λDE3) ,IPTG诱导表达 ,SDS -PAGE以及Western -Blot分析。结果　获得pET32a -ipaC重组表达质粒 ,SDS -PAGE以及Western -Blot显示重组质粒表达产物的分子量约为 6 3kDa,并具有一定的免疫活性。结论　福氏志贺氏菌毒力基因ipaC可在大肠杆菌中表达 ,且表达产物具有一定的免疫活性。Aim To construct a recombinant plasmid encoding Shigella flexneri ipaC gene and detect the immunocompetence of the expressed product in E.coli.Methods Clone the ipaC gene into the downstream of the T7 promoter pET32a,transform the recombinant plasmid into competent cells of E.coli BL21(λDE3).To assess the potential immunocompetence of the recombinant expressed product,induce by IPTG,and analyse with SDS-PAGE and Western-blot.Results The recombinant plasmid pET32a-ipaC SDS-PAGE and Western-Blot indicated the molecular weight of expressed product was about 63kDa and had immunocompetence.Conclusions The recombinant plasmid coding for ipaC gene can express in E.coli BL21(λDE3) and the expressed product have immunocompetence which may be used as the candidate vaccine for Shigella flexneri.

关 键 词：福氏志贺氏菌 毒力基因 IPAC 大肠杆菌 基因表达 细菌性痢疾 

分 类 号：R378.25[医药卫生—病原生物学] R394[医药卫生—基础医学]