引起免疫失败的传染性腔上囊病病毒VP2基因在大肠杆菌中的表达  

Expression in Escherichia coli of VP2 gene of infectious bursal disease virus isolated from chickens with immunoprophylaxis defeat

作　　者：曹冰玉[1,2] 王永山[1] 范红结[2] 欧阳伟[1] 王忠灿[3] 唐雨德[3] 

机构地区：[1]江苏省农业科学院兽医研究所农业部动物疫病诊断与免疫重点开放实验室国家兽用生物制品工程技术研究中心,江苏南京210014 [2]南京农业大学动物医学院,江苏南京210095 [3]南京军区军事医学研究所,江苏南京210002

出　　处：《中国兽医科学》2010年第2期130-134,共5页Chinese Veterinary Science

基　　金：国家自然科学基金项目(30571371);江苏省自然科学基金项目(BK2008352;BK2009041);江苏省农业科技创新基金项目[CX(08)106]

摘　　要：将从安徽省传染性腔上囊病(IBD)免疫失败的病鸡腔上囊组织中扩增的VP2基因定向克隆入原核表达载体pGEX-4T-1中,构建了原核表达质粒pGEX-VP2,并转化大肠杆菌Rosetta(DE3);经IPTG诱导后,用IBDV双抗体夹心ELISA检测,结果呈阳性反应;SDS-PAGE分析结果显示,重组VP2融合蛋白的分子质量为68ku,表达量约占菌体总蛋白量的16.0%,表达产物主要以包涵体形式存在于菌体内;Western-blotting结果显示,重组VP2蛋白与IBDV抗体发生反应,形成一条特异性蛋白带,表明获得的重组蛋白具有良好的抗原反应性。A VP2 gene of infectious bursal disease virus（IBDV） was amplified from bursa of Fabricius of chickens with immunoprophylaxis defeat in Anhui Province and cloned into the prokaryotic expression vector pGEX-4T-1.The expression plasmid pGEX-VP2 was constructed and transformed into competent Escherichia coli Rosetta（DE3）,then the target protein was expressed under the induction with IPTG and was positive in the detection of IBDV sandwich ELISA.SDS-PAGE analysis showed that the VP2 fusion protein was approximately 68 ku in molecular weight,and it made up 16.0% of the total bacterial proteins as inclusion bodies in E.coli.Western-blotting showed that the VP2 protein could react with IBDV antibody,indicating that the VP2 protein possessed good antigenicity.

关 键 词：传染性腔上囊病病毒 VP2基因 原核表达 

分 类 号：S852.659.4[农业科学—基础兽医学]