Construction of eukaryotic expression vector of fusion protein pEGFP/hVEGF165 and its expression in vascular endothelial cells / 中南大学学报(医学版)
Journal of Central South University(Medical Sciences)
; (12): 1047-1050, 2007.
Article
em Zh
| WPRIM
| ID: wpr-813952
Biblioteca responsável:
WPRO
ABSTRACT
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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes de Fusão
/
RNA Mensageiro
/
Transfecção
/
Expressão Gênica
/
Células Cultivadas
/
Células Endoteliais
/
Fator A de Crescimento do Endotélio Vascular
/
Proteínas de Fluorescência Verde
/
Vetores Genéticos
Limite:
Humans
Idioma:
Zh
Revista:
Journal of Central South University(Medical Sciences)
Ano de publicação:
2007
Tipo de documento:
Article