结核分枝杆菌三种基因重组质粒的构建及原核表达  

Construction of recombinant plasmid of three kind of gene from mycobacterium tuberculosis and its prokaryotic expression

作　　者：丁淑琴[1] 师志云[2] 董辉[2] 杨风琴[1] 朱佳佳[1] 

机构地区：[1]宁夏医科大学检验学院,宁夏银川750004 [2]宁夏医科大学总医院医学实验中心,宁夏银川750004

出　　处：《检验医学与临床》2013年第4期388-389,391,共3页Laboratory Medicine and Clinic

基　　金：宁夏自然基金项目(编号:NZ1196);宁夏医科大学校级项目(编号:XM200907;XM201012)

摘　　要：目的构建结核分枝杆菌ESAT-6、CFP10、phoS2/pET28a表达重组质粒,并原核表达重组蛋白。方法将目的基因亚克隆到pET-28a表达载体,转化入大肠埃希菌BL21(DE3)plysS,诱导表达重组蛋白ESAT-6、CFP10、phoS2。结果成功构建基因工程菌株ESAT-6、CFP10、phoS2/pET-28a/BL21(DE3)plysS,表达重组蛋白ESAT-6、phoS2,因以包涵体形式存在,无法纯化。结论成功构建结核分枝杆菌ESAT-6、CFP10、phoS2/pET28a重组质粒,分别表达分子量约10×103、31×103的ESAT-6、phoS2重组蛋白,但无法纯化,CFP10不表达。Objective To construct a recombinant plasmid of ESAT-6,CFP10,phoS2 from mycobacterium tuberculosis,and study on prokaryotic expressing of recombinant protein.Methods Target gene was subcloned into expression vector pET-28a,then transferred into escherichia coli BL21（DE3）plysS,and inducible expressed the recombinant protein of ESAT-6,CFP10,phoS2.Results We constructed genetic engineering strain ESAT-6,CFP10,phoS2/pET28a of Mycobacterium tuberculosis and expressed fusion protein ESAT-6,CFP10 and phoS2.But the fusion protein could not be purifying because they were inclusion bodies.Conclusion The recombinant plasmid ESAT-6,CFP10,phoS2/ pET28a of Mycobacterium tuberculosis were successfully constructed and expressed.The proteins were about and 10×103 and 31×103.Because the fusion proteins are inclusion bodies so they cannot be purified.

关 键 词：结核分枝杆菌 ESAT-6 CFP10 phoS2 重组质粒 原核表达 

分 类 号：R382.31[医药卫生—医学寄生虫学]