Construction and expression of eukaryotic expression vector for hepatitis B virus X gene containing EGFP / 重庆医科大学学报

ABSTRACT

Objective:To construct eukaryotic expression vector for hepatitis B virus X(HBV X) gene with enhanced green fluorescence protein(EGFP),and confirm its expression in hepatocellular carcinoma(HCC)cell line.Method: HBV X gene was cloned from pcDNA3.1(+)-X by enzyme digestion and inserted to pEGFP-N1.The rector was confirmed by enzyme digestion,PCR assay and sequencing.Then pEGFP-X was transfected to HCC cell line Bel-7402.After transient transfection,expressions of HBV X and EGFP gene were detected by RT-PCR and fluorescence microscope respectively.Results: The results showed that the recombinant plasmid could express HBV X gene efficiently in Bel-7402,which showed green fluorescence.Conclution The pEGFP-X was constructed and the fused HBV X-EGFP gene was expressed in Bel-7402 successfully.These facilitate the study of the effect of HBV X protein on the development of HCC.