Expression of green fluorescent protein gene in human nucleus pulposus cells mediated by lentiviral vector

Objective To construct the Lenti-GFP carrying the green fluorescent protein(GFP) gene and to investigate the expression of GFP in human nucleus pulposus cells.Methods The GFP gene was inserted to lentiviral vector plasmid pLenti6/ V5 TOPO by molecular cloning technique.Human kidney 293 cells were co-transfected with the 3 plasmid system containing packaging plasmid and envelope plasmid by using calcium phosphate precipitation,and the expression of GFP was observed under fluorescence microscope after 48 h.The viral particles were collected 72 h after transfection,and were used to infect human nucleus pulposus cells.The expression of GFP in nucleus pulposus cells was observed by fluorescence microscopy.Results The results showed that the transfection efficacy was evident and the viral titer was 107 U /mL.Conclusion The lentiviral vector containing GFP gene is successfully constructed,and the transduction and expression of GFP gene is obvious in nucleus pulposus cells.