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Journal of Central South University(Medical Sciences) ; (12): 1047-1050, 2007.
Artículo en Chino | WPRIM | ID: wpr-813952

RESUMEN

OBJECTIVE@#To construct the plasmid of human vascular endothelial cell growth factor165 and green fluorescence protein report gene eukaryotic expression vector of fusion protein pEGFP /hVEGF165, and to detect its expression in vascular endothelial cells.@*METHODS@#We amplified full-length of gene VEGF165 by PCR, cloned in direction in multiple clone sites of pEGFP-N1, constructed recombinant plasmid of pEGFP/hVEGF165. Through enzyme digestion, PCR, and sequencing analysis, we also performed liposome-mediated transfection of vascular endothelial cells of in vitro cultivation, and detected the expression of fusion protein pEGFP/hVEGF165 using fluorescence microscope, RT-PCR, and Western blot.@*RESULTS@#Both gene VEGF165 and multiple clone site of pEGFP-N1 confirmed by PCR, enzyme digestion, and sequence analysis. EGFP/VEGF protein was expressed in vascular endothelial cells after pEGFP/VEGF165 recombinant plasmid transfected vascular endothelial cells.@*CONCLUSION@#Fusion protein eukaryotic plasmid of report gene EGFP and VEGF165 is successfully constructed, and EGFP/VEGF can be expressed in vascular endothelial cells, which lays a foundation for the application of VEGF gene in treating ischemia vascular diseases.


Asunto(s)
Humanos , Células Cultivadas , Células Endoteliales , Metabolismo , Expresión Génica , Vectores Genéticos , Proteínas Fluorescentes Verdes , Genética , Plásmidos , ARN Mensajero , Proteínas Recombinantes de Fusión , Genética , Metabolismo , Transfección , Factor A de Crecimiento Endotelial Vascular , Genética , Metabolismo
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