| Objective: To construct a cell line conferring resistance toHygromycin and stably expressing hepatitis B e antigen (HBeAg) andhepatitis B surface antigen (HBsAg).Methods: The plasmid pSEH-Flag-HBV was identified by doubledigestion and sequencing, and the hepatoma cell line HepG2cells weretransfected with this plasmid by liposome, and then screened withHygromycin. ELISA. PCR, RT-PCR, Western blotting and Southernblotting were used to detect the expression of HBV in this cell line.Results: The size of the recombinant plasmid pSEH-Flag-HBV wereexpressed and integrated in HepG2, which demonstrated by ELISA, PCR,RT-PCR, Western blotting and Southern blotting.Conclusion: One stable cell line HepG2-H4that HBV genome wasintegrated singly in this cellular DNA is successfully established. |
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