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Progress in Modern Biomedicine. 2014, (11)

Breeding and Identifying Presenilin-1 and Presenilin-2 Conditional Double Knockout MiceChinese Full Text

ZHANG Yuan;GUO Qing-xi;LIU Yong;TANG Hong;PU Xia;RUAN Si-bei;LUO Xia;TANG Ming-xi;Department of Pathology,the Affiliated Hospital of Luzhou Medical College;

Abstract: 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 from all 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.
Keywords：

PS1,PS2; Conditional knockout; Mice; Cre,genotyping,breeding,colony;

  • DOI：

    10.13241/j.cnki.pmb.2014.11.012

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  • Subject：

  • Classification Code：

    Q78

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