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.
出处
《检验医学与临床》
CAS
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)
