牦牛病毒性腹泻病毒E2基因的克隆鉴定及原核表达  

Cloning and Prokaryotic Expression of E2 Genes of Bovine Viral DiarrheaVirus Strain from Yak

作　　者：孙凯[1] 刘亚刚[1] 王妍[1] 王盼盼[1] 胡炳峰[1] 王文伯[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《四川动物》2011年第6期886-889,共4页Sichuan Journal of Zoology

基　　金：四川省科技厅攻关项目(04JY029-006-04);国家民委项目(07XN03)

摘　　要：运用聚合酶链式反应,以牦牛BVDV基因组DNA为模板扩增出牦牛BVDVE2基因。为研究E2蛋白的抗原性,将E2基因插入到pET-32a原核表达载体,构建重组表达质粒pET-32a-E2,并转化至BL21(DE3)宿主菌中,利用IPTG诱导表达。经SDS-PAGE检测,pET-32a-E2在宿主菌BL21(DE3)中表达出约58kD的融合蛋白,结果表明E2基因在BL21(DE3)宿主菌获得了高效表达,表达蛋白大小与预期相符。Western-blotting结果显示E2蛋白具有良好抗原性。这对以后研究E2蛋白功能等具有十分重要的意义。E2 gene was successfully amplified by PCR using genome DNA of the wild yak as template.In order to study the antigenicity of E2 protein,we inserted E2 gene into pET-32a vector and reconstructed plasmid pET-32a-E2 and then transformed them into BL21（DE3） host bacterium,using IPTG to induce them to express in the end.The result showed that a 58 kD fusion protein which was detected by SDS-PAGE was expressed efficiently by pET-32a-E2 in the BL21（DE3） host bacterium and the size of expressed protein was consistent with our anticipation.Western-blotting results showed that E2 protein has a good antigenicity.The research is important for the future study of the function of E2 protein.

关 键 词：牦牛 E2基因 牛病毒性腹泻病毒 原核表达 

分 类 号：S852.65[农业科学—基础兽医学]