血管内皮生长因子 (VEGF1 83)转基因小鼠的建立(英文)

采用显微注射法 ,将一个由 1.3kb人感光细胞间维生素A类结合蛋白 (hIRBP)启动子区和人血管内皮生长因子 (hVEGF183)cDNA所构成的融合基因片段HIRV183导入 2 89枚ICR小鼠受精卵的雄性原核 .制备当代 (G0 )转基因小鼠 ,共得G0 代小鼠 17只 .经PCR和PCR Southern方法筛选得到 1只为hVEGF183转基因阳性小鼠 ,整合率为 5 9%,转基因总效率为 0 3%.将其与正常ICR小鼠交配 ,经PCR和dot blot杂交鉴定后代 (F1)小鼠阳性整合率为 4 5 9%.F1代兄妹之间进行交配 ,后代 (F2 )小鼠阳性整合率为 6 7 6 %.结果表明 ,外源基因hVEGF183整合在小鼠染色体的单一位点 ,为以后利用转基因动物的方法对VEGF这个新型异构体VEGF183进行深入研究打下了基础

Transgenic Mice Carrying the Gene of VEGF183

The transgenic mice were produced by microinjecting a chimeric gene HIRV183 into the male pronucleus of 289 fertilized egg of ICR mice. The HIRV183 was composed of the human interphotoreceptor retinoid binding protein promoter fused to the cDNA fragmen encoding human vascular endothelial growth factor(VEGF183). Out of 17 mice, 1 mouse (G 0) carrying the transgene was identified by PCR and PCR Southern analysis. Total integration rate and efficiency of transgene were 5.9% and 0.3%, respectively. The transgenes were transmitted to the next generation with a ratio of 45.9%, and 67.6% of offspring from F1 carried the transgene. The result indicates that the transgene was integrated into chromosome. This preliminary result supports the feasibility of using transgenic mice to study the biological activity of the long isoform, VEGF183, in the tissue\|specific applications.