Gene Construction Of Adenoviral Vector-Mediated Vascular Endothelial Growth Factor 121

Objective The purpose of this study was to construct the recombinant Adenoviral vector(rAD) expressing human Vascular Endothelial Growth Factor gene-121(VEGF121),and support the study of angiogenesis effect of VEGF121 gene on vascular endothelial cells in vitro and extro.Methods 1.The human VEGF gene was obtained by reverse transcription-polymerase chain reaction ( RT-PCR ) and directly cloned into the pShuttle -IRES-GFP,the recombinant plasmid was called rAD-VEGF121. 2. To genenate the pAD-VEGF121 viral particles, using GFP as a report gene, AD-293 cells was co-transfected with the three plasmids pAD- VEGF121 by the way of liposome-mediated transfection.Results 1. The recombinant AD expressing human VEGF121 gene was successfully constructed and confirmed by DNA sequencing. 2. The recombinant AD was successfully constructed in AD-293 cells.Conclusion The recombinant AD expressing human VEGF121 gene was successfully constructed and this study can be the theoretical basis for the further study of the ischemia diseases in extremity .