Prokaryotic expression and biological functions of Mycobacterium tuberculosis Rv1884c and Rv0867c genes
- VernacularTitle:结核分枝杆菌Rv1884c和Rv0867c基因的克隆表达及其促生长作用的研究
- Author:
Xiaopeng GAO
;
Mingquan SU
;
Jiayun LIU
;
Qiaohong YUE
;
Liu YANG
;
Jianfang ZHANG
;
Xiaoke HAO
- Publication Type:Journal Article
- Keywords:
Myeobacterium tuberculosis;
Bacterial proteins;
Recombinant fusion proteins;
Cytokines;
Gene expression
- From:
Chinese Journal of Laboratory Medicine
2008;31(12):1390-1395
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct fusion gene and prokaryofie expression plasmid encoding the Rv1884c and Rv0867c genes in Mycobacterium tuberculosis (M.tb).The fusion protein wsg expressed efficienfly in E.coli cells.Methods The Rv1884c and Rv0867c genes were amplified by polymerase chain reactions (PCR) with specific primers from genomic DNA of M.tb H37Rv strain,and cloned into pGEX-4Tland pUC19 vectors.Rv1884c and Rv0867c were subcloned into the expression vector pGEX-4T-1 and pPRO-EXHT followed by DNA sequencing.The plasmids were transformed into E.coli DH5α and induced to produce GST-fused Rv1884c and His-fused Rv0867c fusion protein.The protein molecular weight and expression format was analyzed by SDS-PAGE.Results The recombinant expressive vectors pGEX-4T-1Rv1884c and pPRO-EXHT-Rv0867c were constructed.The DH5α strains of E.coli with recombinant plasmid showed high level of Rv1884c and Rv0867c gene expressions after IPTG induction.The SDS-PAGE showed that the plasmids expressed Rv1884c and Rv0867c fusion proteins with molecule weight of 45 000 and 80 000.The recombinant protein accounted for 18.3%and 23.7%of total bacteria protein.The expressed proteins could be purified via GSTrao FF and Ni2+-NTA system kits in denatured condition.Conclusions Mycobacterium tuberculosis Rv1884e and Rv0867c genes have been cloned and expressed Successfully in E.coli DH5α.The results lay a basis for further study of fast culfivation in Mycobacterium tuberculosis and investigation of their activities and functions.
