Construction of cell models transfected with HBV adr subtype

  Objective   To establish HBV replication cell models.

  Methods   Using molecular subclone technique,pcDNA3-3 HBV was constructed by insertion of 9 600 bp fragment of HBV adr subtype genome into the EcoRⅠ and HindⅢ sites of pcDNA3.Hep G2 was transfected with pcDNA3-3 HBV by using lipofectamine transfection reagent and screened with antibiotic G418.

  Results   The plasmid pcDNA3-3 HBV was constructed successfully.After stable transfection,HBsAg and HBeAg were positive in the supernatant which tested by ELISA.S gene and pre-S gene were also positive which tested by PCR.cccDNA was found in the cells by PCR,and HBV pre2S1 mRNA was measured by RT-PCR.

  Conclusion   pcDNA3-3 HBV can be expressed,transcribed and replicated in Hep G2 cells.