空肠弯曲菌外膜蛋白rOMP18的克隆表达及抗原性分析  

Cloning,expression and antigenicity analysis of the outer membrane protein 18 fromCampylobacter jejuni

作　　者：乔博[1,2] 孟凡亮[2] 顾一心[2] 何利华[2] 张姝[2] 肖迪[2] 刘国栋[2] 曹芳芳[2] 张建中[2] 张茂俊[2] 

机构地区：[1]郑州大学公共卫生学院,郑州4500011 [2]中国疾病预防控制中心传染病所,北京102206

出　　处：《中国人兽共患病学报》2011年第1期41-44,共4页Chinese Journal of Zoonoses

基　　金：中国疾病预防控制中心青年基金(2009A102);艾滋病和病毒性肝炎等重大传染病防治(2008ZX10004-002);病原体网络化监测技术研究(2008ZX10004-008)

摘　　要：目的构建空肠弯曲菌omp18基因的重组表达质粒、克隆表达rOMP18重组蛋白、对表达产物进行鉴定并评价其抗原性。方法 PCR扩增空肠弯曲菌的omp18基因片段,将其连接到表达载体pET32a(+)中并转化至大肠杆菌(E.co-liBL21(DE3)),诱导表达后SDS-PAGE分析其表达产物并纯化目的蛋白。应用空肠弯曲菌感染免疫血清,利用Western-blot方法分析其抗原性。结果与结论单、双酶鉴定结果显示已成功构建rOMP18的重组表达载体pET32a-omp18,IPTG诱导表达产物的相对分子质量与理论相符34kD(HIS16kD)。Western-blot检测结果表明重组蛋白rOMP18具有抗原性,为空肠弯曲菌感染的特异抗体的检测提供候选抗原。In order to clone,express and identify the antigenicity of the outer membrane protein rOMP18 ofCampy-lobacter jejuni（C.jejuni）,omp18 was amplified by PCR and ligated to plasmid pET32a（＋）.After the recombinant plasmid was transformed toE.coliBL21（DE3）,the expression was induced by IPTG.The expressed protein was identified with SDS-PAGE and its antigenicity was determined by Western blot（WB） analysis.Restriction enzyme analysis showed the recombinant plasmid pET32a-omp18 have been successfully constructed.The recombinant expressed protein had a weight of 34kD（HIS16kD）,which was similar to the predicted his-tag and the protein rOMP18.WB result indicated the expressed rOMP18 had good antigenicity which inferred it might be a potential antigen for the serological diagnosis ofC.jejuniinfection.

关 键 词：空肠弯曲菌 克隆 表达 rOPM18 抗原性 

分 类 号：R378[医药卫生—病原生物学]