转人组织蛋白酶K基因小鼠的Southern杂交鉴定  

Identification of Transgenic Mice of Cathepsin-K by Southern Blot Hybridization

作　　者：祝梅香[1] 邹星[2] 沈洁[1] 王嫣[1] 徐凤[1] 郭金微[1] 张晶[2] 李亦平[3] 

机构地区：[1]中国医学科学院中国协和医科大学实验动物研究所,北京100021 [2]北京师范大学 [3]美国哈佛大学

出　　处：《中国比较医学杂志》2005年第1期40-42,共3页Chinese Journal of Comparative Medicine

摘　　要：目的　用地高辛标记探针的方法进行Southern杂交 ,以检测转人组织蛋白酶K基因小鼠外源DNA的整合情况 ,筛选携带目的基因的阳性小鼠。方法　用地高辛标记探针的方法 ,对转人组织蛋白酶K基因小鼠 (F0代 )及首建阳性鼠与野生型ICR小鼠交配产生的F1代进行Southern杂交 ,对F0代和F1代进行筛选。结果　经显微注射法 ,共获得 2 5只转基因小鼠 ,通过Southern杂交鉴定 ,1只为阳性 ,阳性率为 4 % ;对PCR阳性的F1代小鼠 35只进行Southern杂交 ,结果全部为阳性。结论　外源基因 (人CathepsinK基因 )已整合到小鼠的染色体上 ,并且能够遗传。Objective Southern blot hybridization with DIG labeling probe was used to identify whether transgenic mice of human Cathepsin-K gene were positive. Methods Transgenic mice and the intercrossed progeny between the founder mice of human Cathepsin-K gene and wild ICR were detected by Southern blot hybridization with DIG labeling probe. Results 25 transgenic mice were obtained through microinjection, one was detected to be positive by Southern blot hybridization. The positive rate is 4%. Positive F1 generation via PCR was all positive by Southern blot. Conclusion The foreign gene (human Cathepsin K gene) has already integrated into mice chromosome and can inherit to the F1 generation.

关 键 词：小鼠 阳性 SOUTHERN杂交 组织蛋白酶 目的基因 地高辛标记探针 FD 外源基因 交配 外源DNA 

分 类 号：R512[医药卫生—内科学] R383[医药卫生—临床医学]