结核分枝杆菌CFP10-ESAT-6融合基因的表达及其多克隆抗体的制备  

Expression of CFP10-ESAT-6 fusion gene of Mycobacterial tuberculosis and preparation of polyclonal antibody against the expressed protein

作　　者：唐宇龙[1] 刘湘新[1] 杨华[2] 丁元生[2] 胡忠义[2] 

机构地区：[1]湖南农业大学动物医学院,湖南长沙410128 [2]上海市结核重点实验室,上海200433

出　　处：《中国兽医科学》2006年第10期811-814,共4页Chinese Veterinary Science

摘　　要：以结核分枝杆菌标准株H37Rv的基因组DNA为模板,经重叠PCR扩增获得CFP10-ESAT-6融合基因片段,克隆于pET21a表达载体中,获得了重组质粒pET21a-CFP10-ESAT-6,将其转化大肠杆菌BL21细胞,经IPTG诱导表达,得到了25 ku的目的蛋白。表达产物经纯化和定量分析后免疫新西兰大白兔,收集兔血清进行抗体ELISA检测。结果显示,成功构建了CFP10-ESAT-6融合基因的原核表达质粒,获得了CFP10-ESAT-6融合蛋白,以此蛋白免疫动物可产生高效价的抗体。The gene encoding protein CFP10-ESAT-6 was amplified from Mycobacterium tuberculosis H37Rv chromosomal DNA by PCR. The PCR product was cloned into pET21a and recombinant pET21a-CFP10-ESAT-6 was transformed into E. coli BL21. The target protein of 25 ku was produced by induction of IPTG. The protein was injected into New Zealand rabbits after it was purified and analysed quantitatively. Then sera from the vaccinated rabbits were analysed by ELISA. ELISA result showed that high titer polyclonal antibody against CFP10-ESAT-6 fusion protein was obtained from the rabbits immunized with the protein.

关 键 词：结核分枝杆菌 融合蛋白CFP10-ESAT-6 表达 多克隆抗体 

分 类 号：S852.618[农业科学—基础兽医学] Q786[农业科学—兽医学]