Breeding of APPSWE transgenic mice and gene identification of filial generation

AIM: To explore the ideal breeding way of APPSWE transgenic mouse (Tg2576) and identify filial generation mice with APPSWE gene for laying a foundation of AD research. METHODS: (1) Three different copulation ways were adopted to observe the survival rate and positive rate with APPSWE gene of filial generation mice. (2) The genome DNA was extracted from the tails of filial generation mice and PCR method was employed to amplify the APPSWE gene fragment. The gene type was observed by electrophoresis.(3) PCR product was inserted in pGEM-T vector to detect its gene sequence. RESULTS: The male Tg2576 mice mated with female Tg2576, and four litters were reproduced, all of them died during two days. The male Tg2576 mice mated with female C57BL, and the survival rate of their filial generation was 81% and positive rate with APPSWE gene was 43.3%. The male C57BL mice mated with the female Tg2576,and the survival rate and positive rate with APPSWE gene of filial generation mice were 82.4% and 21.9%, respectively. By Wilcoxon's Rank Sum Test, there were no significant different between the survival rates of filial generation mice by two breeding way (P＞0.05), but there were significant different between the positive rates with APPSWE gene (P＜0.05). Electrophoresis result showed the molecular weight of PCR product was 428 bp and was in accordance with that of APPSWE gene fragment. By gene sequencing, gene sequence of PCR product was verified as same as APPSWE gene double mutant. CONCLUSION: It is ideal way of breeding filial generation mice with APPSWE gene that male Tg2576 mouse mate with female C57BL. PCR technique can identify the filial generation with APPSWE gene precisely, and offer an ideal animal model for further research on AD.