Construction of reporter gene vector containing two promoters and its expressing activity assay in eukaryotic cells

Objective To study the expressing activity of vector containing two promoters in eukaryotic cells.Methods Reporter gene expressing vectors pC LacZ and pCS LacZ containing CMV promoter and CMV SV40 promoters were constructed respectively.β galactosidase activity transfected with these two vectors in TJ905 cells were assayed.The vectors constructed were transfected into TJ905 cells with cationic liposome,lipofectamine.Preparation of extract from the cells 1 to 3 days post transfection and β galactosidase activity assays were performed.The enzyme activity of the cell extract transfected with standard pSV gal vector was compared with that of transfected with the two vectors constructed.Results The activity of β galactosidase with pCS LacZ was significantly higher than that with the two other vectors( P 0.05).The peak of the enzyme activity occurred 48 hours post transfection.Conclusion This will provide a beneficial exploring way in promoting gene expressing efficiency in futrue gene therapy.