结核分枝杆菌_____结核分枝杆菌；克隆；表达；纯化

结核分枝杆菌Rv2450基因的克隆、表达及纯化

http://www.100md.com 《第四军医大学学报》 2004年第19期

结核分枝杆菌,,结核分枝杆菌；克隆；表达；纯化,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Cloning of Mycobacterium tuberculosis Rv2450 gene and purification of its fusion protein using affinity chromatography

    XUE Ying， JIANG Hong， GAO Xue， BAI YingLan, SHI ChangHong， ZHANG Hai， XU ZhiKai， LI Yuan

    Department of Microbiology， School of Basic Medicine， Fourth Military Medical University， Xi’an 710033， China

    【Abstract】 AIM: To obtain Rv2450 gene segments of Mycobacterium tuberculosis (Mtb), express them efficiently in E.coli and purify the aim proteins. METHODS: Rv2450 gene segments were amplified by PCR with specific primers from genome of Mtb H37Rv strain. After sequenced, Rv2450 gene segments were subcloned into expression vector pProEX HT and purified in E.coli DH5α. Recombinant 6×His fused proteins were purified via NiNTA purification system. RESULTS: The length of PCR products was 529 bp and identical with what the GenBank reported. SDSPAGE analysis showed that the fusion protein mainly existed in inclusion bodies. We isolated the inclusion bodies from the culture and purified the fusion protein under denaturing condition using metal chelate affinity chromatography. CONCLUSION: The construction of the recombinant expressing plasmid lays a basis for further study of Rv2450 protein. ......

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