Construction of the recombinant adenovirus hu-VEGF_(121)

Objective:To construct the adenoviral vector that expresses hu-VEGF 121 gene for further study of the gene therapy on the osteonecrosis.Methods: The hu-VEGF 121 gene was cloned to the shuttle plasmid Track-CMV, the shuttle plasmids were linearized with Pme1 and was transfected to BJ5183 cells which obtain adenoviral backbone vector. The candidate clones were further tested by nuclease digestion and PCR. The recombinant adenoviral plasmids were linearized and transfected to 293 cells, viral production were monitored by GFP(green fluorescent protein)expression. The expression of VEGF 121 was tested by immunohistochemistry. Results:We successfully cloned the hu-VEGF 121 to the shuttle plasmid Track-CMV. The correct recombinant adenoviral plasmid was constructed and recombinant adenovirus hu-VEGF 121 was obtained by transfecting 293 cells.Conclusion:We have constructed the recombinant adenovirus hu-VEGF 121 ,it will provides basis for the further study of gene therapy on the osteonecrosis.