西尼罗河病毒E蛋白结构域Ⅲ基因的原核表达  

Prokaryotic Expression of West Nile Virus E Protein Domain Ⅲ

作　　者：魏艳鸣[1] 朱碧波[1] 曹胜波[1] 

机构地区：[1]华中农业大学动物医学院,武汉430070

出　　处：《湖北农业科学》2012年第12期2593-2595,共3页Hubei Agricultural Sciences

基　　金：国家公益性行业(农业)科研专项(200903037)

摘　　要：根据已发表的西尼罗河病毒(WNV)的DNA序列设计合成一对特异性引物,用聚合酶链式反应(PCR)扩增E蛋白结构域Ⅲ(E-DⅢ)的基因。将PCR产物克隆至pET28a原核表达载体上,获得重组表达质粒pET28a-E-DⅢ。将该质粒转化BL21(DE3),经IPTG诱导后提取包涵体,纯化融合蛋白,通过SDS-PAGE和Western blot方法证实E-DⅢ的基因可以在大肠杆菌中高效表达。A pair of primers was designed according to the reported WNV DNA sequences and applied for amplifying the E-DⅢ fragment by polymerase chain reaction（PCR）.The PCR product was cloned into the pET28a vector.Then the constructed recombinant plasmid pET28a-E-DⅢ was transformed into E.coli BL21（DE3） and induced by IPTG.According to SDS-PAGE and western blot,E-DⅢ protein was expressed at high level.

关 键 词：西尼罗河病毒 E蛋白结构域Ⅲ 原核表达 

分 类 号：S852.659.6[农业科学—基础兽医学] Q786[农业科学—兽医学]