鹿结核分枝杆菌MPB70蛋白的表达与免疫原性鉴定  

Expression of MPB70 Protein and Identification the Immunogenicity of Deer Mycobacterium tuberculosis

作　　者：王晓龙[1] 卢天成[1] 王秀然[1] 杨艳玲[2] 

机构地区：[1]吉林农业大学生命科学学院,长春130118 [2]中国农业科学院特产研究所,长春130112

出　　处：《中国畜牧兽医》2016年第12期3135-3140,共6页China Animal Husbandry & Veterinary Medicine

基　　金：吉林省科技攻关项目(20140204070NY)

摘　　要：本研究以分离的鹿结核分枝杆菌DNA为模板扩增免疫原性蛋白MPB70基因,获得约590bp片段,并将其克隆,构建原核表达载体pET-30a-MPB70,将重组质粒转入大肠杆菌BL21(DE3),经IPTG诱导后纯化和SDSPAGE分析,在20ku处可见特异性蛋白条带。利用鹿结核阳性血清进行Western blotting鉴定,原核表达的融合蛋白可与鹿结核阳性血清抗体结合,并出现特异的免疫反应。该蛋白可作为特异性抗原进行鹿结核病的检测,从而为鹿结核病诊断方法的研究奠定基础。In this study,the immunogenicity protein MPB 70 gene was amplified from Mycobacte-rium tuberculosis genome DNA which separated from deer,and about 590 bp fragment was ob-tained.Then the fragment was cloned and constructed prokaryotic expression vector of pET-30a-MPB70,and the recombinant plasmid was put into E .coli BL21 （DE3）.Purified after IPTG in-duction,and analyzed by SDS-PAGE,a specificity protein band was observed at 20 ku.Using the deer serum positive of tuberculosis in Western blotting,the fusion protein could be combined with deer serum positive of tuberculosis antibody and arise specific immune response.The protein could be used as a specific antigen to test the deer tuberculosis.The study laid a foundation for further studying the deer tuberculosis appraisal method.

关 键 词：结核分枝杆菌 MPB 70 基因 原核表达 

分 类 号：S852.618[农业科学—基础兽医学]