人转铁蛋白转基因克隆牛整合位点检测  

Detection of the Integration of Human Transferrin Gene on Chromosomes in Transgenic Cloned Cows

作　　者：刘宇[1] 朱怡文[1] 许淼[1] 黄英[1] 

机构地区：[1]上海市儿童医院上海医学遗传研究所,卫生部医学胚胎与分子生物学重点实验室暨上海市胚胎与生殖工程重点实验室,上海200040

出　　处：《中国畜牧兽医》2012年第2期60-63,共4页China Animal Husbandry & Veterinary Medicine

基　　金：国家“转基因生物新品种培育”科技重大专项(2009ZX08007-003B)

摘　　要：为检测外源基因整合在转基因克隆牛染色体上的定位情况,通过优化染色体荧光原位杂交技术,成功对4头人转铁蛋白转基因克隆牛的整合位点进行定位。每头牛分析中期分裂相40个以上,杂交率在53.1%～86.8%之间,且杂交信号分别位于转基因克隆牛的11号、15号和19号染色体的相应位置;另外采用定量PCR方法对杂交信号强弱不同的转基因克隆牛个体进行整合位点拷贝数的检测,结果显示拷贝数的多少与杂交信号强弱直接有关。To detect the integration of human transferrin gene（hTF） on chromosomes in transgenic cloned cows,the author optimized fluorescence in situ hybridization（FISH）.For transgenic cloned cows,53.1% to 86.8% of metaphases showed hybridization signals which were located on different chromosomes,such as Chr.11,Chr.15 and Chr.19.The results showed that using FISH to detect the integration site and hybridization signals was highly efficient and specific,also the copy numbers were detected by Real-time PCR.

关 键 词：荧光原位杂交 人转铁蛋白转基因克隆牛 整合位点 拷贝数 

分 类 号：Q813.4[生物学—生物工程]