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.
出处
《中国畜牧兽医》
CAS
北大核心
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)
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
fluorescence in situ hybridization
human transferring transgenic cloned cows
integration site
copy number
