转人组织蛋白酶K基因小鼠的Southern杂交鉴定

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(FD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25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Cathepsin K基因)已整合到小鼠的染色体上，并且能够遗传。

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

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.