Package of recombinant retrovirus and detection of titre of hepatitis B virus (adr hypotype)

Objective To establish a basement for constructing human hepatitis B virus(adr) transgenic mice model with packaging and determining titer of hepatitis B recombinant retrovirus.Methods The recombinant retroviral vector was obtained by inserted full-length HBV genome(adr)in retroviral vector pLNCL. Following identification of forward and reverse direction connection,the vectors were transfected into PA317 packaging cell line by liposome transfection.Two kinds of cells were screened with G418.G418-resistant PA317 colonies were amplified and concentrated to detect the viral titers.Results Two kinds of hepatitis B recombinant retrovirus were yielded.The titer of one virus with forward direction connection was 10~6 CFU/ml,the titer of another virus was 10~5 CFU/ml. Conclusions The recombinant retroviral with full-length HBV genome(adr) was packed by PA317 cell.