Construction and expression of Retroviral Vector containing HBx gene / 重庆医科大学学报

ABSTRACT

Objective:To construct the retroviral vector containing HBx gene so as to investigate the effects of HBx gene in the pathogenesis of HBV-dependent hepatic cell carcinoma(HCC)and further construct animal models of hepatic cell carcinoma.Methods:HBx gene was amplified from the HBx adenovirus Plasmid by polymerase chain reaction(PCR)technique whose primer was designed according to the announced sequences of HBx gene in gene bank,then cut by two endonucleases and directionally cloned into the pseb-hus vector and analyzed by gene sequencing analyzer,the correct constructed vectors were transfected into L02 cell line.At last,HBx protein was detected by Western Blotting.Results:Through PCR,endonuclease cutting and gene sequencing,the target gene was verified into be correctly cloned to retroviral plasmid pSEB-HUS and the high expression of green fluorescence protein(GFP)in L02 cell line was observed under fluorescent microscope.The expression of HBx protein was also observed by Western Blotting.Conclusion:Recombinant retroviral vector expressing HBx protein was successfully constructed.