牛分支杆菌ag85b基因的克隆及表达  

Cloning and Prokaryotic Expression of ag85b Gene of Mycobacterium Bovis

作　　者：马玢[1,2] 曾瑾[1,2] 李武[1,2] 刘晓明[1,2] 邓光存[1,2] 

机构地区：[1]宁夏大学西部特色生物资源保护与利用教育部重点实验室,宁夏银川750021 [2]宁夏大学生命科学学院,宁夏银川750021

出　　处：《动物医学进展》2013年第5期44-47,共4页Progress In Veterinary Medicine

基　　金：国家自然科学基金项目(31160515)

摘　　要：以牛分支杆菌C68001株基因组DNA为模板,克隆免疫优势抗原基因ag85b,构建重组表达质粒pET-28a-ag85b,转化入大肠埃希菌BL21(DE3),IPTG诱导表达重组蛋白,对表达产物进行SDS-PAGE分析和Western blot鉴定。结果表明,构建了ag85b基因的重组基因工程菌株,IPTG诱导表达的重组蛋白分子质量约为33ku,目的蛋白能被His单克隆抗体识别,表达产物主要以包涵体形式存在。To clone and express the geneen coding Mycobacterium tuberculosis antigen Ag85B, a pair of spe- cific primers were designed, the Ag85B gene was amplified from the genomic DNA by PCR, and cloned in- to vector PET28a （＋） to construct the recombinant plasmid. The recombinant plasmid was transformed into the expression vector Escherichia coli BL21 （DE3）, and induced with IPTG; the presence of the recombinant protein in the expression vector was analysed by SDS-PAGE and Western blot. Results showed that the ag85b gene was successfully expressed with expected 33 ku in molecular weight. It can be specifically recognized by His McAb.

关 键 词：牛分支杆菌 Ag85B抗原 原核表达 

分 类 号：S852.618[农业科学—基础兽医学]