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.