Construction and application of retroviral vector system useful for high-efficiency transfection into HepG2 cells

To construct the retroviral expression vector carrying green fluorescent protein gene(EGFP) and human insulin gene and study expression of the vector in HepG2 cells.The fragment encoding IRES-EGFP was cloned into the retroviral expression vector carrying human insulin gene with regulatory element(pLXSN-GI-Ins).The expression plasmid of pLXSN-GI-Ins-EGFP were constructed.The plasmid was transfected into HepG2 cells by using lipofectamine.To observe the expression of EGFP gene by fluorescence microscope cells cultured in medium with glucose at concentration of 30.0 mmol/L.Our results suggest that the retroviral expression plasmid of pLXSN-GI-Ins-EGFP was successfully constructed and the expression of EGFP gene was(38.0±5.0)%.Thus,the retroviral expression vector carrying green fluorescent protein gene(EGFP) and human insulin gene with regulatory element was successfully constructed and expressed in HepG2 cells.