Comparision of the efficiency of different internal promoters driving the expression of greeen fluorescent protein mediated by lentiviral vector

Objective Three kinds of different internal promoters were inserted into the lentiviral vector deriving from human immunodeficiency virus-1(HIV-1) to drive the expression of the green fluorescent protein(GFP) and the efficiencies of the internal promoters were compared.Methods The restriction enzemies and T_4 DNA ligase were used to construct the vector plasmids,and the recombinant plasmids were identified by the restriction exzemies.Human embroic kideny 293T cells were cotransfected with the three plasmids by calcium phosphate DNA precipation,and the expression of GFP was observed under fluorescent microscope.The titers of the lentiviral vectors were determinde by scoring GFP expression following swerial dilutions of the viral supernatant on 293T cells.Results The plasmids containing the PPT element and different internal promoters and GFP gene were constructed.The transfected 293T cells were found containing strong expression of GFP.The amount of the 293T cells expressing GFP driven by CMV promoter was largest among all promoters,while the LSP promoter was the smallest.The titer of the lentiviral vector with CMV internal promoter was 5×10~6IU/ml and others were 1～2×10~5IU/ml.Conclusion Under our experiment condition,the efficiency of CMV promoter was the strongest,the LSP promoter was the weakest,while the PUB promoter was at somewhere between the CMV and the LSP promoters.