[Construction of prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II and expression in E. coli].

OBJECTIVE

To Construct the prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II.

METHODS

IFN-alpha1b 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-alpha1b was constructed. Circumsporozoite protein II (CSP II) 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-alpha1b was cut from the recombinant plasmid pGEX-4T-1/IFN-alpha1b digested with BamH I and EcoR I and ligated with the recombinant plasmid pGEX-4T-1/CSP II also digested with BamH I and EcoR I. The recombinant prokaryotic plasmid pGEX-4T-1/IFN-alpha1b/CSP II was constructed. The fusion gene IFN-alpha1b/ CSP II was expressed in E. coli by IPTG.

RESULTS

The prokaryotic expression vector pGEX-4T-1/IFN-alpha1b, pGEX-4T-1/CSP II and pGEX-4T-1/IFN-alpha1b/CSP II were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-alpha1b/CSP I in E. coli was identified by SDS-PAGE and Western blot.

CONCLUSION

The prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II was successfully constructed, which was then expressed in E. coli.