Construction of hVEGFA gene recombinant adenovirus vector and its efficiency of transfection

Objective: To construct the recombinant adenovirus vector containing hVEGFA gene.Methods: The hVEGFA gene coding sequence was cloned into pAdTrack-CMV plasmid to get pAdTrack-CMV-hVEGFA,which would be transformed into competent E.coli BJ5183 carried backbone plasmid pAdeasy-1 already.The homologous recombinant adenoviral plasmid was delivered into HEK293A cells after verification.The identified recombinant adenovirus(rAd-hVEGFA) was amplified in HEK293A cells.Viral particle concentration was determined by TCID50.Results: The recombinant adenovirus vector was constructed successfully.The expression of GFP was observed and the right gene was found.Conclusion: The recombinant adenoviral vector carrying hVEGFA gene was successfully constructed,and provided the basis of ischemic cerebrovascular disease′s gene therapy.