牛病毒性腹泻病毒牦牛分离株E0基因的克隆表达  

Cloning and Expression of E0 Gene of Bovine Viral Diarrhea Virus from Yak

作　　者：叶成玉[1] 王光华[1] 蔡其刚[1] 王戈平[1] 陆艳[1] 张芳芳 马利青[1] 周继章 

机构地区：[1]青海省畜牧兽医科学院,青海西宁810016 [2]家畜疫病病原生物学国家重点实验室农业部兽医公共卫生重点开放实验室,甘肃兰州730046

出　　处：《动物医学进展》2012年第9期16-20,共5页Progress In Veterinary Medicine

基　　金：科技部基础研究专项(2008FY210200)

摘　　要：将青海牦牛牛病毒性腹泻病毒(BVDV)青海泽库(QHZK)株的E0基因亚克隆入原核表达载体pET-32(a),构建了重组表达载体pET-32(a)-E0,然后用重组质粒转化Rosetta(DE3)感受态细胞,并利用IPTG诱导蛋白表达。表达的蛋白用His-Band镍柱进行亲合层析纯化,Western blot鉴定表达蛋白。结果显示,E0基因可在大肠埃希菌中获得表达,表达产物的分子质量约为44ku,与预期的蛋白分子质量大小一致;Western blot分析表明,该蛋白可以与BVDV标准阳性血清产生特异性结合反应。To express and purify the recombinant protein of the E0 of bovine viral diarrhea virus（BVDV）,the E0 gene of BVDV QHZK strain was cloned into pET32a（＋） vector.The recombinant plasmid was transformed into Rosetta（DE3） competent cells,and was induced with IPTG.The expressed products were analyzed by SDS-PAGE and Western-blot,and purified using affinity chromatography.The results showed that the E0 gene was expressed in E.coli and the expressed protein was 44 ku in size,as it was expected.Western blot test demonstrated that the expressed protein could specifically react with BVDV standard antiserum.The results showed that the expressed protein with satisfactory antigencity,laid the foundation for the development of BVDV subunit vaccines.

关 键 词：牛病毒性腹泻病毒 E0基因 原核表达 抗原性 

分 类 号：S852.659.6[农业科学—基础兽医学]