[Construction and expression of dual promoter lentiviral vector containing TFR and VEGF genes]

To construct a dual promoter lentivira vector containing TFR and VEGF gene, and detect the expression of TFR and VEGF genes in MSCs of Chinese mini-swine.

The TFR and VEGF gene were amplified by polymerase chain reaction and cloned into pLenti-GFP-Neo after the CMV and SV40 promoter respectively, to generate the lentivira vector pLenti-TG-VEGF. The four-plasmids lentiviral vector system(pRsv-REV, pMDlg-pRRE, pMD2G and pLenti-TG-VEGF)were cotransfected into human embryonic kidney 293 T cells with Lipofectin 2000 reagent. The packaged virus was harvested 72 h later. MSCs were infected by lentivirus carrying TFR and VEGF genes. Western blotting was used to detect the expression of TFR and VEGF genes in the viral infected MSCs.

Restriction endonuclease digestion analysis and DNA sequencing demonstrated that the dual promoter lentivirus vector containing TFR and VEGF genes were constructed successfully. MSCs could be successfully infected by the recombinant lentivirus. Expression of TFR and VEGF protein could be detected in the infected MSCs by Western blotting.

The dual promoter lentiviral vector containing TFR and VEGF genes are constructed successfully which provides basis for future research on Cardiac molecular imaging of cell transplantation.