Expression of HBsAg with Three Kinds of Eukaryotic Expression Plasmids Inserted HBsAg Gene in EBV-Immortalized B-cell Line

Objective To investigate the expression of HBsAg with three kinds of eukaryotic expression plasmids inserted HBsAg gene in Epstein-Barr virus-immortalized B-cell line. Methods Three kinds of plasmids inserted HBsAg gene were constructed (pCI-S?pMEP4-S? pLXSN-S) and their accuracy was confirmed by restriction enzyme digestion. Then these recombinant plasmids were transfected into EBV-immortalized B-cells by means of cationic liposome,respectively.Cells were selected with G418 or hygromycin B,and mRNA of HBsAg gene in these selected cells were detected by RT-PCR and HBsAg in culture supernatants and cell lysates were assayed with enzyme-linked immunosorbent assay(ELISA)kit. Results mRNA of HBsAg in selected cells transfected three kinds of eukaryotic expression plasmids inserted HBsAg gene all can been detected by RT-PCR, and these transfected cells produce HBsAg which is readily detectable in culture supernatants and cell lysates.But level of HBsAg expressived by pMEP4-S in EBV-immortalized B-cells is higher than those expressived by pCI-S or pLXSN-S. Conclusions Transfection of EBV-immortalized B-cell line with three kinds of plasmids inserted HBsAg gene all lead to stable expression of HBsAg. Level of HBsAg expressived by pMEP4-S in EBV-immortalized B-cells is higher than those expressived by pCI-S or pLXSN-S .