[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.


CONCLUSIONS
The prokaryotic expression vector of the fusion gene IFN-alpha1b/CSP II was successfully constructed, which was then expressed in E. coli.
Chen
Hui-hong
HH
Department of Parasitology, Health and Science Center of Peking University, Beijing 100083, China.
Yu
Xin-bing
XB
Gao
Xing-zheng
XZ
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005822:Genetic Vectors; D016898:Interferon-alpha / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*
D016898:Interferon-alpha; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; C041595:circumsporozoite protein, Protozoan