文献类型：期刊文献

中文题名：猪瘟病毒E2蛋白抗原表位集中区的原核表达及鉴定

英文题名：Prokaryotic Expression and Identification of the Main Antigenic Domain in E2 Glycoprotein of Classical Swine Fever Virus

作者：郭东光[1];朱艳平[2];银梅[1];宁红梅[1];潘耀谦[1];刘卫[1];陈明艳[1];王选年[1,2]

第一作者：郭东光

机构：[1]河南科技学院;[2]新乡学院生物技术研究中心

第一机构：河南科技学院,河南新乡453003

年份：2013

期号：3

起止页码：15-20

中文期刊名：动物医学进展

外文期刊名：Progress in Veterinary Medicine

收录：CSTPCD;;北大核心:【北大核心2011】；CSCD:【CSCD_E2013_2014】；

基金：河南省基础与前沿技术研究(122300410003);河南省自然科学基金项目(330002;122300410003);国家自然科学基金(31201877);新乡学院博士科研启动经费(1399020164);河南科技学院研究生创新基金项目

语种：中文

中文关键词：猪瘟病毒;E2蛋白;抗原表位;原核表达;鉴定

外文关键词：CSFV ; E2 glycoprotein; antigenic epitope ; prokaryotic expression ;identification

摘要：为获得猪瘟病毒(CSFV)E2蛋白抗原表位集中区原核重组目的蛋白,建立CSFV抗原和抗体快速检测方法。通过扩增CSFV E2蛋白抗原表位集中区基因,亚克隆至表达载pET-30a(+),构建重组原核表达载体pET-30a-E2-1,转化至宿主菌Rostta(DE3)后进行诱导表达。SDS-PAGE分析显示,出现与目的蛋白大小一致的蛋白条带,分子质量大小为31ku。对重组蛋白表达条件进行优化,最终获得其诱导表达的最佳温度为18℃,最佳IPTG浓度为0.1mmol/L。目的蛋白以包涵体形式存在,其表达量约占菌体总蛋白含量的46%。Western blot分析显示,目的蛋白不仅能与兔抗CSFV高免血清反应,还可以与载体特异性标签His单抗反应,表明融合蛋白具有良好的抗原性,为CSFV抗原和抗体检测试剂盒的研制奠定了基础。
In order to gain the main antigen domain of the classical swine fever virus （CSFV） E2 protein, to establish rapid detection method for CSFV antigen and antibody. The main antigenic domain of CSFV E2 was amplified and subcoloned into prokaryotic expression plasmid pET-30a（＋） vector. The recombinant plasmid named pET-30a-E2-1 was transformed into E. coli Rosette （DE3） induced by IPTG. Recombinant expression products were analyzed by SDS-PAGE, a specific expression band in form of inclusion body with a relative molecular weight 31.0 ku was detected. The optimum temperature was 18℃, the IPTG was 0.1 mmol/L. The expression amount accountsed for about 46% of the yield in total cell lysate. Western blot indicated that the expressed proteins were recognized not only by antibody against the C-strain of CSFV , but also by the specific monoclonal antibody of HIS tag. The result suggests that the recombinant protein has good antigenicity and provides substantial base for the CSFV antibody and antigen detection kit research.