Abstract: Objective To Construct the prokaryotic expression vector of the fusion gene IFN-α1b/CSPⅡ. MethodsIFN-α1b 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-α1b 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-α1b was cut from the recombinant plasmid pGEX-4T-1/IFN-α1b 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-α1b/CSPⅡ was constructed. The fusion gene IFN-α1b/CSPⅡ was expressed in E.coli by IPTG. Results The prokaryotic expression vector pGEX-4T-1/IFN-α1b, pGEX-4T-1/CSPⅡ and pGEX-4T-1/IFN-α1b/CSPⅡ were identified by PCR, enzyme digestion and gene sequencing. The expressed fusion protein/IFN-α1b/CSPⅡ in E.coli was identified by SDS-PAGE and Western blot. Conclusion The prokaryotic expression vector of the fusion gene IFN-α1b/CSPⅡ was successfully constructed, which was then expressed in E.coli .

Key words: Fusion gene IFN-α1b/CSPⅡ, Circumsporozoite proteinⅡ(CSPⅡ), Recombinant DNA, Sequence analysis, Gene expression