Construction, package and identification of retroviral vector of androgen recetor antisense RNA

To construct, pack and identify the retroviral vector of androgen receptor (AR) antisense RNA (pL-AR-SN). PCR amplified fragment of AR cDNA was inserted inversely into the retroviral vector pLXSN. PA317 cells were transfected by the recombinant retroviral vector with liposome. The genomic DNA of the transfected PA317 cells and the retrovirus RNA from the supernatant of cell culture were extracted and identified by PCR and RT-PCR. NIH 3T3 cells were used to determine the virus titer of the supernatant. pL-AR-SN was proved to contain the inversely inserted fragment of AR cDNA by restriction endonuclease digestion. The virus titer of the supernatant was 2.34X10~(5) CFU/ml. The inserted fragment could be amplified by both PCR and RT-PCR from the genomic DNA and the retrovirus RNA. The retroviral vector of androgen receptor antisense RNA has been constructed successfully and the transfected PA317 cells can produce high titer recombinant virus.