Construction and expression of eukaryotic expression vectors of HBV antigen genes

Objective:To analyze the expression of HBV antigen genes in eukaryotic cells in vitro,eukaryotic expression plasmids containing HBV antigen genes were constructed.Methods:The fragments of HBV antigen genes were amplified by PCR from the plasmid PcDNA3.1-HBV which contains 1.3 fold HBV whole genome sequence.Then the PCR fragments were cloned into Peasy-T1-Simple vector following the routine procedures.After identification by enzyme digestion and DNA sequence analysis,the interested gene fragments were inserted into the eukaryotic expression vector PcDNA3.1(+).The resultant recombinant plasmids were confirmed by restriction enzyme digestion and the correct ones were transiently transfected into HepG2 cells,and LO2 cells as a control.Western blot and Immunofluores-cence were used to analyze the expression of HBV gene products in the transfected cells.Results:The fragments of HBV antigen genes were correctly amplified.Restriction enzyme digestion and DNA sequence analysis confirmed that the recombinant plasmids containing HBV antigen genes were successfully constructed.Western blot and Immunofluorescence confirmed that the interested genes could be correctly expressed in transfected cells.Conclusion:The construction of eukaryotic expression plasmids containing HBV antigen genes and the antigen gene expression in eukaryotic cells in vitro provide a solid experimental foundation for further research on the biological function of HBV antigens.