Construction and identification of a recombinant lentiviral expression vector carrying on Med19 gene and its expression in 293T cells

Objective To construct a lentiviral expression vector of human Med19 and identify its expression in 293T cells.Methods Human Med19 sequence was amplified,purified,ligated with lentiviral vector plasmid and verified by sequencing.The verified plasmids were transfected into 293T cells by Lipofectamine 2000. Transfection efficiency was assayed by both immunofluorescence microscopy and western blot.Medl9 lentiviral vector plasmid from the selected constructs was propagated and harvested with a virus packaging system,and the virus titers were determined.Results The Medl9 gene was successfully constructed into pGC-FU-Med19 express lentiviral vector.Medl9 expression was observed using fluorescence microscope after the transfection.Western blotting also showed Medl9 expression in the transfected 293T cells.The Med19 gene sequence of the vector was successfully verified by sequencing.The concentrated titer of virus suspension was 2×10~9 Tu/mL.Conclusion The human Medl9 lentiviral expression vector was successfully constructed.This facilitates further studying Medl9 biological function.