【摘要】 目的 :探讨老年性痴呆 (AD)转基因模型鼠 (TgAPPSWE2 576杂合子鼠 ,Tg2 576)的优化繁育方法 ,鉴定APPSWE基因阳性的子代鼠 ,为下一步AD研究奠定基础。方法 :(1)用 3种不同的交配方式观察子代鼠的存活率及APPSWE基因阳性的比率 ;(2 )从子鼠鼠尾中提取基因组DNA ,用PCR方法扩增APPSWE基因片段 ,电泳后观察结果 ;(3 )PCR产物装入pGEM -Teasy载体中 ,测序证实其基因序列。结果 :雄性和雌性Tg2 576互交 ,所产 4窝子鼠 ,2d内全部死亡。雄性Tg2 576和雌性C57BL交配所产子鼠存活率 81% ,APPSWE基因的阳性率为 43 3 %。雌性Tg2 576和雄性C57BL交配存活率 82 4% ,APPSWE基因的阳性率 2 1 9%。经秩和检验两种繁育方式所产子鼠的存活率无显著差异 (P >0 0 5) ,APPSWE基因的阳性率有显著差异 (P <0 0 5)。琼脂糖电泳显示PCR产物分子量为 42 8bp ,与目的基因片段相对分子质量大小一致 ;DNA序列测定证实PCR产物基因序列和APP片段基因序列一致 ,并存在APPSWE(K670N ,M671L)突变。结论 :雄性Tg2 576和雌性C57BL品系小鼠交配是繁育APPSWE基因阳性子鼠的较好方法 ;本实验所用PCR方法能够精确鉴定APPSWE基因阳性子鼠 ,为AD的实验研究提供了较理想的动物模型更多还原

【Abstract】 AIM: To explore the ideal breeding way of A PPSWE transgenic mouse (Tg2576) and identify filial generation mice with APP SWE ge ne for laying a foundation of AD research. METHODS: (1) Three di fferent copulation ways were adopted to observe the survival rate and positive r ate with APPSWE gene of filial generation mice. (2) The genome DNA was extra cted 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. RESU LTS: The male Tg2576 mice mated with female Tg2576, and four litters wer e 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 posit ive rate with APPSWE gene was 43 3%. The male C57BL mice mated with the fem ale Tg2576,and the survival rate and positive rate with APPSWE gene of filial g ener ation mice were 82 4% and 21 9%, respectively. By Wilcoxon’s Rank Sum Test, th ere were no significant different between the survival rates of filial generatio n mice by two breeding way (P>0 05), but there were significant differen t between the positive rates with APPSWE gene (P<0 05). Electrophoresi s re sult 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 prod uct 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 gene ration with APPSWE gene precisely, and offer an ideal animal model for furth er research on AD.更多还原