The Construction of Eukaryotic Expression Vector of HBV x Gene

Objective This study was designed to construct the eukaryotic expression vector of HBV X gene for exploring the contribution of X gene to the chronic hepatitis and the hepatocellular. Methods X gene with EcoRⅠand HindⅢ endoenzyme sites was obtained by using PCR. Vector PUC118 and X gene was digested with double enzyme. Both fragments were connected using ligase and transferred to Eco-JM109 and named as PUC118-HBx , double enzyme digestion was conducted for vector PUC118-HBx and PCDNA3.1(+), both fragments were recovered and connected and transferred to Eco-DH5α Reconstituted plasmid sequence was examined by auto-sequencing assay and named as PCDNA3.1(+)-HBx. Results We have constructed the eukaryotic expression vector of X gene that carry integra X gene fragment by using auto-sequencing assay. Conclusion PCDNA3.1(+)-HBx eukaryotic expression vector is helpful in studying the role of HBV X gene and X protein in carcinogenesis of chronic hepatitis B virus and hepatocellular carcinoma, and provide a gene material for studying the biological functions of HBV X gene and X protein.