| 张元 郭庆喜 刘勇 唐红 蒲霞 阮思蓓 罗霞 唐明希.Presenilin-1/Presenilin-2双基因敲除小鼠的繁育及基因型鉴定[J].现代生物医学进展英文版,2014,14(11):2052-2054. |
| Presenilin-1/Presenilin-2双基因敲除小鼠的繁育及基因型鉴定 |
| Breeding and Identifying Presenilin-1 and Presenilin-2 ConditionalDouble Knockout Mice |
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| DOI： |
| 中文关键词: presenilins 基因 基因敲除 小鼠 聚合酶链反应 琼脂糖凝胶电泳 |
| 英文关键词: PS1, PS2 Conditional knockout Mice Cre, genotyping, breeding, colony |
| 基金项目:四川省科技厅科技支撑计划(2012SZ0172) |
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| 中文摘要: |
| 目的：繁殖及鉴定Presenilins双基因敲除小鼠,为进一步研究阿尔茨海默症(AD)奠定基础。方法：将引进的野生型及PS1/PS2
双基因敲除小鼠进行饲养并繁殖，繁殖成功的子代小鼠基因型有野生型、杂合子和纯合子3种。提取子代小鼠鼠尾基因组DNA，
用PCR 法和琼脂糖凝胶电泳鉴定基因类型。结果：PS1/PS2 双基因敲除小鼠的饲养和繁殖均获得成功，繁殖结果符合孟德尔遗传
规律，同时获得更多基因型小鼠和Presenilins 双基因敲除小鼠。结论：正确的饲养繁殖以及鉴定方法是获得PS1/PS2 双基因敲除
小鼠的有效途径。 |
| 英文摘要: |
| Objective:To breed and identify presenilin-1 (PS1) and presenilin-2 (PS2) conditional double knockout mice (dKO),
in order to further study the molecular and neuronal mechanisms underlying the pathogenesis of Alzheimer's disease (AD). Methods:Based on the genotypes of the initial breeders that were transported into China from our collaborator at USA, four mating pairs were set
up. The plug was confirmed at about 3-5 days after breeding, and plugged female mice were carefully maintained in the animal facility at
the College. After delivery, pups were weaned at postnatal day 20 (P20), and a small fragment of tail was clipped from each pup. The
genomic DNA was extracted, and PCR with three sets of primers that respectively recognize and distinguish the Cre transgene, PS1
floxed/wild-type alleles, and PS2 knockout/wild-type alleles was performed. The amplified DNA fragments were separated by agarose gel
electrophoresis, and visualized by UV. Results:PCR products fromall three PCR amplifications showed the correct size for each allele as
it was expected. Based on the distributions of genotypes among the pups, the penetration of each genotype was accordant with the
Mendelian inheritance law. At this stage, we have obtained PS1/PS2 homozygous dKO mice. Conclusion:The introduced dko mice from
USA have been successfully bred in our animal facility, and the colonies are under expanding currently. |
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