Construction and validation of a hepatitis B virus replication and expression vector

Objective To construct a HBV replication and expression vector,and then verify its replication and expression in HepG2 cells.Methods The HBV replication and expression vector was constructed by molecular techniques such as PCR,cloning,restriction digestion.The HBV vector was transiently transfected into HepG2 cells with Fugene HD transfection Regent.The HBsAg,HBeAg and HBV DNA secreted into culture medium were detected at different time points.Results Restricted enzymes digestion and sequencing results confirmed that the HBV replication and expression vector was successfully constructed.When the vector was transiently transfected into HepG2 cells,the HBsAg,HBeAg and HBV DNA could be detected in the supernatant of transfected cells.Conclusion A replication and expression HBV vector has been successfully constructed via molecular cloning techniques,which could replicate and be expressed in HepG2 cells.By further modification,it is expected to be an useful tool for HBV phenotypic resistance and replication activity examination.