文献类型：期刊文献

中文题名：鸡传染性支气管炎病毒S1基因抗原区的原核表达及鉴定

英文题名：Prokaryotic Expression and Identification of the S1 Gene Main Antigen Domain of Avian Infectious Bronchitis Virus

作者：朱艳平[1,2];郭东光[1];岳锋[1,2];杨媛[3];李冲[3];王爱国[3];李博文[3];阮涛[3];孔令芸[3];王选年[1]

第一作者：朱艳平

机构：[1]新乡学院生物技术研究中心;[2]新乡学院生命科学与技术系;[3]河南科技学院动物科学学院

第一机构：新乡学院生命科学技术学院

年份：2014

卷号：41

期号：6

起止页码：55-59

中文期刊名：中国畜牧兽医

外文期刊名：China Animal Husbandry & Veterinary Medicine

收录：CSTPCD;;北大核心:【北大核心2011】；

基金：河南省基础与前沿技术研究(122300410003;330002;122300410150);国家自然科学基金(31201877);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：鸡传染性支气管炎病毒;S1蛋白;原核表达;鉴定

外文关键词：avian infectious bronchitis virus ;S1 protein; prokaryotic expression; identification

摘要：本研究以鸡传染性支气管炎病毒(infectious bronchitis virus,IBV)S1基因为研究对象,设计特异性引物扩增S1抗原集中区目的基因,长度为198bp,亚克隆至原核表达载体pGEX-6P-1,构建重组原核表达质粒pGEX-6P-1-S1,转化至宿主菌Rosetta(DE3)后进行诱导表达。SDS-PAGE分析结果显示,表达获得1条特异性蛋白条带,其分子质量大小为33.0ku,与预期蛋白大小一致。可溶性分析结果表明目的蛋白以包涵体形式存在。Western blotting结果显示,重组蛋白能与IBV鸡阳性高免血清反应,被GST标签单抗识别,结果表明该融合蛋白正确表达并具有良好的抗原性。本研究为制备IBV单克隆抗体奠定了抗原基础。
Avian infectious bronchitis virus （IBV） S1 gene was the object in this study and a pair of primers was designed for amplifying the SI main antigen fragment of IBV, which was composed of 198 bp. And then it was sub-cloned into prokary- otic expression plasmid pGEX-6P-1. The recombinant plasmid pGEX-6P-1-S1 was transformed into E. coli Rosetta （DE3） and induced by IPTG. SDS-PAGE analysis showed a specific protein band with a molecular weight 33.0 ku, according to our study anticipation. The express production was inclusion body when it was identified. Western blotting indicated that the expressed protein could react with the chicken positive serum and be recognized by the specific monoclonal antibody of GST. The result suggested that the recombinant protein was correctly expressed and had a good antigenicity, which provided substantial antigen base for preparation of IBV monoclonal antibody.