人血管内皮细胞生长因子165基因真核表达载体构建  

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

作　　者：吴波[1] 陈秉[2] 熊群英[1] 容振勤 张伟林[1] 李永浩[3] 

机构地区：[1]广东省江门中医药学校,529000 [2]广东药学院 [3]中山大学眼科医院

出　　处：《医学研究杂志》2006年第1期16-17,共2页Journal of Medical Research

基　　金：广东省医学科学技术研究基金资助课题(基金号:WSTJJ20031202413001630109304)

摘　　要：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hVEGF165cDNA,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VGEF165cDNA;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。Objective To clone human vascular endothelial growth factor 165 （hVEGF165）, construct its eukaryotic expression and to study the expression of hVEGF165. Methods Human vascular endothelial growth factor cDNA was amplified by PCR method from the HL60 cells and cloned to expression vector pcDNA3, constructed pCD-hVGEF165 recombinant plasmid, then transformed to E.coli BL21 （DE3） cell. Results The cloned cDNA was confirmed to be VGEF165 cDNA. It was observed that the expression of human VEGF gene was detected distinctly 72h aftot transferring. Conclusions We successfully cloned and expressed hGEF165 gene, which provided the further foundation of the model of VEGF transgenic animal and makes a basis for the further study in retinal neovascularization.

关 键 词：血管内皮生长因子 基因克隆 基因表达 真核表达载体 眼部新生血管性疾病 

分 类 号：R77[医药卫生—眼科] R394[医药卫生—临床医学]