TI:Construction of Prokaryotic Expression Vector of the Fusion Gene IFN2α1b/ CSP Ⅱand Expression in E. coli

AU:CHEN Huihong , YU Xinbing , GAO Xingzheng

AD:( Depa rtment of Pa rasitology , Health and Science Center of Peking University , Beijing 100083 , China)

【Abstract】　Objective 　To Construct the prokaryotic expression vector of the fusion gene IFN2α1b/ CSP Ⅱ. 　Methods IFN2α1b was amplified from the human genomic DNA by PCR and cloned into prokaryotic expression vector p GEX24T21. The recombinant plasmid p GEX24T21/ IFN2α1b was constructed. Circumsporozoite protein Ⅱ(CSP Ⅱ) was amplified from the Plas2 modium f alciparum genomic DNA by PCR and was cloned into the prokaryotic expression vector p GEX24T21. The recombi2 nant plasmid p GEX24T21/ CSP Ⅱwas constructed. IFN2α1b was cut from the recombinant plasmid p GEX24T21/ IFN2α1b di2 gested with Bam H Ⅰand EcoR Ⅰand ligated with the recombinant plasmid p GEX24T21/ CSP Ⅱalso digested with Bam H Ⅰand EcoR Ⅰ. The recombinant prokaryotic plasmid p GEX24T21/ IFN2α1b/ CSP Ⅱwas constructed. The fusion gene IFN2α1b/ CSP Ⅱwas expressed in E. coli by IPTG. 　Results 　The prokaryotic expression vector p GEX24T21/ IFN2α1b , p GEX24T2 1/ CSP Ⅱand p GEX24T21/ IFN2α1b/ CSP Ⅱwere identified by PCR , enzyme digestion and gene sequencing. The expressed fu2 sion protein/ IFN2α1b/ CSP Ⅱin E. coli was identified by SDS2PAGE and Western blot . 　Conclusion 　The prokaryotic ex2 pression vector of the fusion gene IFN2α1b/ CSP Ⅱwas successfully constructed , which was then expressed in E. coli .

【Key words】　Fusion gene IFN2α1b/ CSP Ⅱ; Circumsporozoite protein Ⅱ( CSP Ⅱ) ; Recombinant DNA; Sequence analysis; Gene expression