Construction and expression of the prokaryotic expression vector of MTb lhp-esat6 fusion gene
- VernacularTitle:结核分枝杆菌lhp-esat6融合基因原核表达载体的构建和表达
- Author:
Quan CHEN
- Publication Type:Journal Article
- Keywords:
Mycobacterium tuberculosis(MTb);
Lhp-esat6 fusion gene;
Recombinant CFP10-ESAT6;
Prokaryotic expression
- From:
Journal of Chongqing Medical University
1986;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct lhp-esat6 fusion gene and its prokaryotic expression vector and express it in E.coli.Methods:By Gene SOEing techniques,a fusion gene was constructed by splicing lhp gene and esat6 gene,then cloned into pQE30 plasmid and expressed in DH5a.Results:The fusion gene was identified by DNA sequencing.A fusion protein about 26kDa was expressed in the E.coli.In presence of 1mM IPTG for 4h,the fusion protein was expressed to the maximum.The fusion protein existed in cytoplasm in soluble form and represented about 40% total bacterial protein of E.coli.Its antigenicity was confirmed by Western blotting.The fusion protein was purified through the Ni-NTA resin and the purity reached 98%.Conclusion:The lhp-esat6 fusion gene and the prokaryotic expression vector (pQE30-CFP10-ESAT6) were constructed successfully,and the fusion protein CFP10-ESAT6 was obtained,so that it can provide an experimental basis for application of the recombinant CFP10-ESAT6.
