作　　者： ; ; ; ; ; ;

机构地区： 天津出入境检验检疫局

出　　处： 《中国兽医科学》 2006年第7期523-528,共6页

摘　　要： 采用PCR方法,以牛疱疹病毒Ⅰ型基因组DNA为模板扩增gB基因,克隆至pGEM-T载体。将以克隆质粒pGEM-T-gB为模板扩增的gBⅠ、gBⅡ和gBⅢ基因片段分别连接至原核表达载体pET32a。获得的重组质粒分别转化感受态细胞BL21(DE3),在IPTG诱导下表达。经SDS-PAGE鉴定,gBⅠ、gBⅡ和gBⅢ基因片段在大肠埃希氏菌BL21(DE3)中以融合蛋白形式获得了表达。以磁化组氨酸蛋白纯化系统对融合蛋白纯化后,Western-blotting和ELISA分析结果表明,这些融合蛋白均与BHV-1标准阳性血清反应,可作为抗原用于牛传染性鼻气管炎ELISA检测方法的建立。 The gB gene was amplified from genomic DNA of bovine herpesvirus-1 by PCR and was cloned into pGEM-T vector. Three overlapping DNA fragments of gB Ⅰ , gB Ⅱ and gB Ⅲ amplified from pGEM-T-gB were ligated with pET32a, and the resultant expression vectors were transformed into E. coli BL21（DE3） cells. The fusion proteins of His-gB Ⅰ ,His-gBⅡ and His-gBⅢ were successfully expressed in E. coli BL21（DE3） and were determined by SDS-PAGE. Western-blotting and ELISA results demonstrated that the fusion proteins purified by magneHis protein purification system could react with positive serum against BHV-1, indicating that these fusion proteins are an antigen source for the development of ELISA for the detection of infectious bovine rhinotracheitis.

关 键 词： 牛疱疹病毒 型 基因 原核表达 纯化 抗原性分析

领　　域： [农业科学] [农业科学] [农业科学]