Construction of Prokaryotic Expression Vector of the Fusion Gene IFN-?1b/CSPⅡ and Expression in E.coli
- VernacularTitle:融合基因IFN-?1b/CSPⅡ原核表达载体的构建及在大肠埃希菌中的表达
- Author:
Huihong CHEN
;
Xinbing YU
;
Xingzheng GAO
- Publication Type:Journal Article
- Keywords:
Fusion gene IFN-?1b/CSPⅡ;
Circumsporozoite proteinⅡ(CSPⅡ);
Recombinant DNA;
Sequence analysis;
Gene expression
- From:
Chinese Journal of Parasitology and Parasitic Diseases
1987;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To Construct the prokaryotic expression vector of the fusion gene IFN-?1b/CSPⅡ. MethodsIFN-?1b was amplified from the human genomic DNA by PCR and cloned into prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/IFN-?1b was constructed. Circumsporozoite proteinⅡ(CSPⅡ) was amplified from the Plasmodium falciparum genomic DNA by PCR and was cloned into the prokaryotic expression vector pGEX-4T-1. The recombinant plasmid pGEX-4T-1/CSPⅡ was constructed. IFN-?1b was cut from the recombinant plasmid pGEX-4T-1/IFN-?1b digested with BamHⅠ and EcoRⅠ and ligated with the recombinant plasmid pGEX-4T-1/CSPⅡ also digested with BamHⅠ and EcoRⅠ . The recombinant prokaryotic plasmid pGEX-4T-1/IFN-?1b/CSPⅡ was constructed. The fusion gene IFN-?1b/CSPⅡ was expressed in E.coli by IPTG. Results The prokaryotic expression vector pGEX-4T-1/IFN-?1b, pGEX-4T-1/CSPⅡ and pGEX-4T-1/IFN-?1b/CSPⅡ were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-?1b/CSPⅡ in E.coli was identified by SDS-PAGE and Western blot. Conclusion The prokaryotic expression vector of the fusion gene IFN-?1b/CSPⅡ was successfully constructed, which was then expressed in E.coli .
