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2004•Journal of Nantong MedicalRequires access

Construction of recombinant adenovirus coding for vascular endothelial growth factor gene

Ping Cai

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Abstract

Objuctive:To estabilish a recombinant adenovirus vector bringing hVEGF 165 cDNA.Methods:Human vascular endothelial growth factor(h VEGF 165 ) cDNA were cloned into adenovirus shuttle vector pHCMVSP1A by standard procedure; recombinant adenoviral plasmid was identified and then transferred to the adenoviral packaging cell HEK 293 cell by lipofectamine mediated gene transfer method to pack the virus;after verifying ,we enlarged and titled the virus.Results:The recombinant Ad.hVEGF 165 was correctly constructed and confirmed by restrictionendonucleas analysis and DNA sequencing analysis; the virus concentration reach to 2.5×10 9pfu/ml.Conclusion:The recombinant adenoviral vector bringing hVEGF 165 cDNA was successfully established.

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What this paper is about

Objuctive:To estabilish a recombinant adenovirus vector bringing hVEGF 165 cDNA.Methods:Human vascular endothelial growth factor(h VEGF 165 ) cDNA were cloned into adenovirus shuttle vector pHCMVSP1A by standard procedure; recombinant adenoviral plasmid was identified and then transferred to the adenoviral packaging cell HEK 293 cell by lipofectamine mediated gene transfer method to pack the virus;after verifying ,we enlarged and titled the virus.Results:The recombinant Ad.hVEGF 165 was correctly constructed and confirmed by restrictionendonucleas analysis and DNA sequencing analysis; the virus concentration reach to 2.5×10 9pfu/ml.Conclusion:The recombinant adenoviral vector bringing hVEGF 165 cDNA was successfully established.

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Available abstract

Objuctive:To estabilish a recombinant adenovirus vector bringing hVEGF 165 cDNA.Methods:Human vascular endothelial growth factor(h VEGF 165 ) cDNA were cloned into adenovirus shuttle vector pHCMVSP1A by standard procedure; recombinant adenoviral plasmid was identified and then transferred to the adenoviral packaging cell HEK 293 cell by lipofectamine mediated gene transfer method to pack the virus;after verifying ,we enlarged and titled the virus.Results:The recombinant Ad.hVEGF 165 was correctly constructed and confirmed by restrictionendonucleas analysis and DNA sequencing analysis; the virus concentration reach to 2.5×10 9pfu/ml.Conclusion:The recombinant adenoviral vector bringing hVEGF 165 cDNA was successfully established.

Key concepts: Recombinant DNA, Complementary DNA, Lipofectamine, Viral vector, Shuttle vector, Adenoviridae, Vascular endothelial growth factor, Molecular biology

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