文献类型：期刊文献

中文题名：猪瘟病毒E0蛋白的原核表达及鉴定

英文题名：Prokaryotic Expression and Identification of E0 Protein of Classical Swine Fever Virus

作者：郭东光[1];朱艳平[1];孙国鹏[1];岳峰[1];贾文科[1];王军[1,2];李鹏[1,2];王自浩[1];王选年[1]

机构：[1]新乡学院生命科学与技术系生物技术研究中心;[2]郑州大学

第一机构：新乡学院生命科学技术学院

年份：2014

卷号：35

期号：5

起止页码：31-35

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项目(122300410003);河南省自然科学基金研究项目(330002)(122300410003);河南省教育厅科学技术研究重点项目(13A230837)

语种：中文

中文关键词：猪瘟病毒;E0;蛋白;鉴定

外文关键词：CSFV;E0 protein;identification

摘要：为获得猪瘟病毒(CSFV)E0重组蛋白,建立CSFV抗体快速检测方法。本研究通过扩增CSFV C株E0基因,亚克隆至原核表达载体pGEX-4T-2,构建重组原核表达质粒pGEX-4T-2-E0,转化至宿主菌Rostta(DE3)后进行诱导表达。SDS-PAGE检测表明与目的蛋白大小一致,其分子质量大小为46ku。优化重组蛋白表达条件,最终获得诱导表达的最佳温度为37℃,最佳IPTG浓度为0.1mmol/L,并且目的蛋白以包涵体形式存在。Western blot表明,目的蛋白能与CSFV兔化高免血清反应,表明该融合蛋白具有良好的抗原性。本研究为CSFV抗体检测试剂盒的研究奠定了基础。
In order to gain the recombinant protein E0 of the classical swine fever virus（CSFV）,to estab-lish CSFV antibody rapid detection method,the CSFV C strain E0 was amplified and sub-cloned into pro-karyotic expression plasmid pGEX-4T-2 vector.The recombinant plasmid named pGEX-4T-2-E0 was transformed into E.coli Rosetta （DE3）and induced by IPTG.The expression products were analyzed by SDS-PAGE,a specific expression band,in form of inclusion body with a molecular weight 46.0 ku was de-tected.The optimum temperature was 18℃,the IPTG was 1.0 mmol/L.Western-blot indicated that the expressed protein was recognized not only by the rabbit antibody to CSFV,but also with the specific mon-oclonal antibody of GST tag.The result suggests that the recombinant protein has good antigenicity and provides substantial base for the CSFV antibody detection kit research.