Expression and purification of truncated C gene combined with preS1 gene from HBV in E.coli

    HU Xingª²Bin, YIN Wen, WEI Sanª²Hua, LEI Yingª²Feng, L¦a Xin, SUN Mengª²Ning, YANG Jing, XU Zhiª²Kai

    Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To construct the prokaryotic recombinant plasmid to express HBV truncated C gene with preS1 gene and to acquire and purify the fused protein for antigenic analysis. METHODS: The target gene was amplified by PCR and digested by restrictive enzyme before cloned into pET28a. It was then transformed into E.coli DH5¦Á and the positive recombinant plasmid named pET28aª²Ctª²preS1 was sequenced. The target protein was distinctly expressed after transformed into E.coli BL21 and induced with IPTG. The protein was scanned and purified on Ni2+ª²NTA column and Western blot was performed after solubility analysis. RESULTS: The recombinant plasmid pET28aª²Ctª²preS1 was successfully constructed and could efficiently express the target gene. Protein production mainly was in inclusion body but could be purified through Ni2+ª²NTA column. The purified protein maintained the antigen activity. CONCLUSION: The truncated HBcAg combined with preS1 antigen can efficiently be expressed and purified, which lays a foundation for further research of novel vaccine against HBV. ......