hVEGFA recombinant adenovirus vector and its transfection efficiency of.docVIP

 PAGE \* MERGEFORMAT 15 hVEGFA recombinant adenovirus vector and its transfection efficiency of Of: INNER, Yuan Zhicheng, Chen Bo, Lu Peisong, Liqiao Yu, Cao Kan [Abstract] Objective: AdEasy replication-defective adenovirus vector system constructed recombinant adenovirus vector carrying hVEGFA. Methods: Molecular cloning, plasmid from PCR2.1 hVEGFA hVEGFA cloned gene coding sequence, digestion, the connection After the system into the adenovirus shuttle vector pAdTrack CMV, the recombinant plasmid pAdTrack CMV hVEGFA, be transformed with adenovirus backbone vector pAdeasy 1 of the Escherichia coli BJ5183, homologous recombination, the recombinant sub pAdeasy 1 hVEGFA. Identification , will pAdeasy 1 hVEGFA transfected human embryonic kidney cells (HEK293A cells), obtained with hVEGFA recombinant adenovirus (rAd hVEGFA). HEK293A cell proliferation repeated adenovirus infection, 50% tissue culture infective dose method (TCID50) detection virus titer, fluorescent microscopy of green fluorescent protein (GFP) expression. Results: We successfully constructed a gene containing hVEGFA adenovirus rAd hVEGFA, HEK293A cells transfected with green fluorescent protein expression after the show that adenovirus rAd hVEGFA transfection successfully received by a large number of repeated virus infections HEK293A cells. CONCLUSION: The constructed gene containing the hVEGFA adenovirus rAd hVEGFA, for ischemic cerebrovascular disease gene vector for gene therapy. [Keywords:] hVEGFA gene; adenovirus vector; transfection; ischemic cerebrovascular disease [Abstract] 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 recombi