鸡传染性支气管炎病毒S1基因抗原区的原核表达及鉴定

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198 bp，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33.0 ku，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。

Prokaryotic Expression and Identification of the S1 Gene Main Antigen Domain of Avian Infectious Bronchitis Virus

Avian infectious bronchitis virus (IBV) S1 gene was the object in this study and a pair of primers was designed for amplifying the S1 main antigen fragment of IBV, which was composed of 198 bp. And then it was sub-cloned into prokaryotic 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.