红系特异的GFP基因在转基因小鼠中的整合和表达3.PDFVIP

动物学报 　50 (2) :263 - 268 , 2004 A cta Zoologica S inica 　　　　　 　 红系特异的 GFP 基因在转基因小鼠中的整合和 表达 颜景斌　肖艳萍　陈美珏　黄淑帧　曾溢滔 上海交通大学医学遗传研究所 , 上海 200040 摘 　要 　应用荧光定量 PCR 技术对由位点控制区 LCR 的 HS2 元件和β 珠蛋白基因启动子指导的红系特异表达 绿色荧光蛋白 ( GFP) 基因的转基因小鼠中外源基因拷贝数进行测定 , 使用荧光显微镜和流式细胞仪检测小鼠 外周血中 GFP 的表达水平 , 并运用荧光原位杂交技术 ( FISH) 确定了其中两只转基因小鼠中外源基因的整合位 点 , 结果表明 : 在转基因小鼠中外源基因的拷贝数各不相同且相差较大 , 而且拷贝数与 GFP 基因的表达量之间 未呈现出相关性 ; FISH 分析确定出两只转基因小鼠的外源基因整合于不同的染色体上 ; 杂交信号的强弱与拷贝 数的多少相一致 [动物学报 50 (2) : 263 - 268 , 2004 ] 。 关键词 　荧光定量 PCR 　荧光原位杂交 　转基因小鼠 　拷贝数 　表达 Integration and expression of GFP gene directed by the erythroidspecif ic element in transgenic mice YAN JingBin , XIAO YanPing , CHEN MeiJ ue , HUAN G ShuZheng , ZEN G YiTao Institute of Medical Genetics , Shanghai Jiaotong University , Shanghai 　200040 , China Abstract 　To study the relationship between integration status and expression level of the foreign gene , cop y number of the foreign green fluorescence protein ( GFP) gene directed by the erythroidspecific elements ( HS2 and HS3) in trans genic mice were determined by realtime quantitative PCR , and the GFP expression level in peripheral blood was measured by fluorescenceactivated cell sorter ( FACS) analysis and was observed under a fluorescence microscope. The integrated sites of foreign gene in two transgenic mice were detected using fluorescene in situ hybridization ( FISH) . The results showed that the copy numbers of the foreign gene were different and vari