Construction of pEGFP-IFI16 Expression Vector and the Influence of Its Expression on Hep-2 Cell Proliferation

IFI16 gene amplified by RT-PCR was ligated into the pUCm-T vector and confirmed by sequencing,and then ligated into the pEGFP-C1 vector to construct the pEGFP-IFI16 recombinant plasmid.The recombinant plasmid pEGFP-IFI16 transfected into Hep-2 cells and its expression was analyzed with fluorescent microscope and semi-quantitative RT-PCR,and the influences of its expression on Hep-2 cell proliferation was further analyzed through assaying cell growth curve using flow cytometry.The results showed that the pEGFP-IFI16 recombinant plasmid was correctively constructed,and after transfecting Hep-2 cells,the green fluorescent signals was observed with fluorescent microscope,and the semi-quantitative RT-PCR result showed that the band brightness of IFI16 gene was obviously increased after transfection in Hep-2 cells,meanwhile it was found from cell growth curve assay that after transfection Hep-2 cell proliferation was slowed down from the second day on and to the third day the cell proliferation rate was obviously slower than that of the control cells.All results illustrated that the pEGFP-IFI16 recombinant plasmid expressing EGFP-IFI16 fusion protein in Hep-2 cells was successfully constructed and its expression could inhibit Hep-2 cell proliferation.