Abstract:

Abstract:Objective  To construct the shRNA lentiviral vectors targeting human NOB1 gene and detect its effect of gene silence in SKOV3 and HEY cells.  Methods The specific siRNA sequences targeting human NOB1 gene were cloned into pLVTHM lentiviral vector. After screening for the valid siRNA,the lentivirus particles were packaged and NOB1 specific shRNA was transmitted into SKOV3 and HEY cells. Then,real-time PCR was performed to determine the expression level of NOB1.  Results  PCR and sequencing results revealed that shRNA plasmids were correctly constructed. The virus with a titer of 8×10⁸ PU/L was successfully packaged.The  NOB1 expressions in SKOV3 and HEY cells were  down-regulated at mRNA level by virus infection. The expression levels of NOB1 mRNA was decreased by 73.5%  in SKOV3 and by 82.2% in HEY cells compared with negative control.

Key words: nin one binding protein;short hairpin RNA;lentivirus