Construction of specifically expressed vascular endothelial growth factor_(165) gene in retina / 中华眼底病杂志

ABSTRACT

Objective To construct specifically expressed vascular endothelial growth factor (VEGF) 165 gene in retina. Methods Rho promoter, specifically expressed in retina, was amplified by polymerase chain reaction (PCR) from the genomic DNA of a BLAB/C rat, then it was cut with restriction enzymes and cloned into the plasmid pcDNA 3.1+-VEGF 165 to form recombinant plasmid pcDNA 3.1+-rho-VEGF 165. The correct recombinant plasmid pcDNA 3.1+-rho-VEGF 165 was identified by restriction enzymes and PCR, and was transferred by jetPEI into cultured human navel vein endothelial cells and human retinal pigment epithelial (RPE) cells. The expression of VEGF protein in human navel vein endothelial and RPE cells was detected by immunocytochemical staining and protraction of the growth curve of the cells. Results In human RPE cells, the expression of VEGF protein was more in recombinant plasmid pcDNA 3.1+-rho-VEGF 165 than that in plasmid pcDNA 3.1+-VEGF 165; in human navel vein endothelial cells, no obvious difference of the expression of VEGF protein between recombinant plasmid pcDNA 3.1+-rho-VEGF 165 and plasmid pcDNA 3.1+-VEGF 165 was found. Conclusions The construction of pcDNA 3.1+-rho-VEGF 165 carrier may provide the basic material for the study of the nosogenesis of VEGF in retinal neovascularization, and establish the foundation to set up the model of transgenic mice with VEGF specific expressing in retina.