Cloning,Expression and Identification of HCV NS5B with C-Terminal 21 aa Truncated

To construct the engineering E.coli.expressing HCV NS5B protein with C-terminal 21 aa truncated and to acquire the protein for screening the anti-HCV drugs targeting the NS5B.The coding region of HCV NS5BC21 gene was amplified by PCR and was digested by BamHⅠand XhoⅠ,the gene fragment was cloned into plasmid pET-28a(+) and the recombinant plasmid was transformed into E.coli BL21(DE3).The positive recombinant plasmid was obtained by identification of PCR amplification,and it was named pET-28a(+)-NS5B-C21. The engineering E.coli.harboring the recombinant pasmid was induced by IPTG and analysed by SDS-PAGE,NS5B-C21 protein was efficiently expressed,and the solubility of expressed NS5B-C21 protein was increased obviously.The expressed NS5B-C21 protein was purified with Ni-Affinity column,and highly pure NS5B-C21 protein was obtained successfully,which established the foundation for screening the anti-HCV drugs targeting the NS5B.