人血管内皮细胞生长因子165基因真核表达载体构建

目的获得人血管内皮生长因子（hVEGF165）cDNA，构建其真核表达载体。方法采用PCR技术，从HL60细胞中扩增出hVEGF165 cDNA，将其克隆至pcDNA3真核表达载体上，构建成为pCD-hVGEF165重组质粒。结果经酶切鉴定，克隆的基因片段为人hVGEF165 cDNA；建立了pCD-hVGEF165重组质粒。结论成功地克隆和表达出了人VEGF165基因，为大量制备重组质粒的研究奠定了基础。

Construction of Eukaryotic Expression Vector Containing Human Vascular Endothelial Growth Factor 165 Gene

Objective To clone human vascular endothelial growth factor 165(hVEGF_ 165 ),construct its eukaryotic expression and to study the expression of hVEGF_ 165 . Methods Human vascular endothelial growth factor cDNA was amplified by PCR method from the HL60 cells and cloned to expression vector pcDNA_3,constructed pCD-hVGEF_ 165 recombinant plasmid, then transformed to E.coli BL21(DE3)cell.Results The cloned cDNA was confirmed to be VGEF_ 165 cDNA. It was observed that the expression of human VEGF gene was detected distinctly 72h after transferring.Conclusions We successfully cloned and expressed hGEF_ 165 gene, which provided the further foundation of the model of VEGF transgenic animal and makes a basis for the further study in retinal neovascularization.