Construction and Expression of a Recombinant Retrovirus Vector Inserted HBsAg Gene

Objective To construct a recombinant retrovirus expression plasmid inserted HBsAg gene and study its expression in eukaryotic cells. Methods The full length gene of HBsAg was obtained from the recombinant plasmid pBKS-S by digesting with EcoRⅠ and BamHⅠ. Then the gene was subcloned into retrovirus expression plasmid pLXSN. The accuracy of recombinant plasmid pLXSN-S was confirmed by restriction enzyme digestion and DNA sequencing;Finally, pLXSN-S was transfected into HepG2、K562 and EBV-immortalized B-cells(EBVC) by means of cationic liposome, respectively. Cells were selected with G418 and HBsAg in culture supernatants were assayed with enzyme-linked immunosorbent assay(ELISA)kit. Results Restriction enzyme digestion and DNA sequencing confirmed that the recombinant retrovirus plasmid inserted HBsAg gene(pLXSN-S)had been constructed correctly and HBsAg were readily detectable in culture supernatants of pLXSN-S transfected cells. Conclusions A retrovirus expression plasmid pLXSN-S has been constructed successfully. Transfection of eukaryotic cells with plasmid pLXSN-S lead to stable expression of HBsAg.