Construction,identification and expression of recombinant Bb-MPT64 vaccine of mycobacterium tuberculosis
- VernacularTitle:结核分枝杆菌重组Bb-MPT64疫苗的构建、鉴定及表达
- Author:
Ting XIE
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis;
Recombinant Bb-MPT64 vaccine;
Construction;
Identification;
Expression
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct and identify recombinant Bb-MPT64 vaccine of mycobacterium tuberculosis. Methods: MPT64 antigen gene were amplified by PCR from mycobacterium tuberculosis H37Rv,then the gene was cloned into Escherichia coli-Bifidobacteria shuttle plasmid pGEX-1?T to construct pGEX-MPT64.The recombinant plasmid was electroporated into Bifidobacteria bifidum(Bb) to construct rBb-MPT64 vaccine.The expression of pGEX-MPT64 was induced with isopropyl-?-D- thiogalactopyranosid(IPTG) and protein MPT64/GST was examined by SDS-PAGE and Western blotting techniques. Results:688bp MPT64 gene was successfully amplified by PCR and cloned into pGEX-1?T by restriction endonuclease analysis.rBb-MPT64 vaccine was successfully constructed by PCR.SDS-PAGE and Western blotting demonstrated that the protein MPT64/GST were expressed in E.coli,BL21. Conclusion:rBb-MPT64 vaccine of mycobacterium tuberculosis is successfully constructed,which may provide basis for the development of new anti-tuberculosis vaccine.
