文献类型：期刊文献

中文题名：猪瘟病毒NS2-3抗原集中区基因的原核表达及鉴定

英文题名：Prokaryotic Expression and Identification of the Main Antigenic Domain Gene in NS2-3 of Classical Swine Fever Virus

作者：刘卫[1];朱艳平[2];宁红梅[1];银梅[1];郭东光[1];鲁毅[1];王选年[1,2]

第一作者：刘卫

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,新乡453003

年份：2013

卷号：32

期号：2

起止页码：165-169

中文期刊名：基因组学与应用生物学

外文期刊名：Genomics and Applied Biology

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

基金：国家自然科学基金(31201877);河南省基础与前沿技术研究(122300410003)(122300410150);河南省教育厅科学技术研究重点项目(13A230837)共同资助

语种：中文

中文关键词：猪瘟病毒;NS2—3;抗原表位;原核表达;免疫印迹

外文关键词：Classical swine fever virus （CSFV）, NS2-3, Antigenic epitope, Prokaryotic expression, Western-blot

摘要：为获得猪瘟病毒(classical swine fever virus,CSFV)NS2-3抗原集中区蛋白,并建立CSFV抗体快速检测方法。本研究以CSFV全长基因组质粒为模板,PCR扩增NS2-3抗原表位集中区,利用扩增片段和克隆载体,构建重组表达质粒,命名为pET32a-NS2-3-1。重组表达质粒转化Rosetta(DE3)细胞,利用IPTG诱导表达,SDS-PAGE电泳和Western-blot鉴定重组表达产物。结果表明,重组质粒pET32a-NS2-3-1在28℃诱导5h得到高效表达,重组蛋白能够与兔抗CSFV阳性血清发生反应。获得CSFVNS2-3抗原集中区蛋白,并且获得的重组蛋白具有抗原性,能够作为CSFV抗体检测的抗原。
To identify the main antigen domain of the classical swine fever virus （CSFV） NS2-3, a method of detecting antibody of classical swine fever virus was established. CSFV genome plasmid as PCR template, the main antigenic domain of CSFV NS2-3 was amplified by PCR, amd PCR products was cloned into prokaryotic expression plasmid pET-32a（＋） vector to obtain the recombinant expression plasmid, named as pET32a-NS2-3-1. The recombinant expression plasmid was transformed into E. coli Rosetta （DE3） and inducible expression by IPTG. The expression products were analyzed by SDS-PAGE and identified by Western-blot. The recombinant plasmid pET32a-NS2-3-1 was highly expressed, induced 5 hours could get a large number of recombinant protein at 28℃, and the recombinant protein reacted strongly with the C-Positive serum of CSFV. The main antigenic domain protein in NS2-3 of CSFV was obtained, which has good antigenicity for detecting antibodies against CSFV.