采用PCR技术,从原核表达质粒PUC19-VEGF165中获得上下游含有Hind Ⅲ和BamH I酶切位点的目的基因VEGF165,与真核载体pEGFP-C1构建重组质粒pEGFP-VEGF165,观察其在内皮细胞中的表达.结果 表明,带有绿色荧光蛋白报告基因的真核表达质粒pEGFP-VEGF165构建成功,并在PEI的介导下成功转染脐静脉内皮细胞(HUVEC),在荧光显微镜下可见强绿色荧光蛋白表达,ELISA检测证明VEGF在细胞上清中有效表达,RT-PCR证明了VEGF165 mRNA水平上的有效表达.此研究结果将为再狭窄的基因治疗提供实验基础.

1. Hutter R.Carrick FE.Valdiviezo C Vascular endothelial growth factor regulates reendothelialization and neointima formation in a mouse model of arterial injury 2004(16)
2. Geiger F.Bertram H.Berger I Vascular endothelial growth factor gene-activated matrix (VEGF165-GAM) enhances osteogenesis and angiogenesis in large segmental bone defects 2005(11)
3. Vera Janavel G.Crottogini A.Cabeza Meckert P Plasmid-mediated VEGF gene transfer induces cardiomyogenesis and reduces myocardial infarct size in sheep 2006(15)
4. Veillette C.Schroeder H Endothelin-1 down-regulates the expression of vascular endothelial growth factor-A associated with osteoprogenitor proliferation and differentiation 2004(7)
5. Ferrara N Molecular and biological properties of vascular endothelial growth factor 1999(7)
6. Yang R.Bunting S.Ko A Substially attenuated hemodynamic response to Escherichia coli-derived vascular endothelial growth factor given by intravenous infusion compared with bolus injection 1998(1)
7. Zhang G.Gurtu V.Kain S R An enhanced green fluorescent protein allows sensitive detection of gene transfer in mammalian cells 1996(3)