Pumilio1/Pumilio2双基因条件性敲除小鼠模型的建立及基因型鉴定

Establishment and Genotyping of Pumilio1 and Pumilio2 Double Conditional Knockout Mouse Strain

目的建立及鉴定Pumilio1/Pumilio2(Pum1和Pum2)双基因条件性敲除的小鼠模型,为进一步研究PUF家族在哺乳动物精子发生中的生物学功能奠定基础。方法将构建的Pum1条件性敲除和Pum2条件性敲除的两个品系小鼠进行交配并繁殖,最终繁殖成功的子代小仔Pum1和Pum2两个基因均为纯合子。对子代以剪趾方式进行编号,提取子代小鼠脚趾或鼠尾基因组DNA,用PCR法和琼脂糖凝胶电泳鉴定基因型。结果 Pum1和Pum2双基因条件性敲除的小鼠模型繁殖成功,同时获得更多PUF家族双基因条件性敲除的小鼠。结论成功建立了Pum1和Pum2双基因条件性敲除的小鼠模型;正确的交配繁殖策略和鉴定方法是获得PUF家族双基因条件性敲除小鼠的有效途径。

Objective To establish and genotype a Pumiliol （Puml） and Pumilio2 （Pure2） double conditional knockout （dCKO） mice, in order to further study the biological function of PUF family in mammalian spermatogenesis. Method Homozygtes of dCKO offspring were produced from crossbreeding of Puml conditional knockout （CKO） mice and Pure2 CKO mice. The pups were identified using toe-marking system at postnatal day 10 （P10） , and weaned at P21. Genomic DNA was extracted, and genotyping was performed using PCR, agarose gel electrophoresis and gel-imaging. Result Puml and Pum2 dCKO mice were obtained and reproduced normally. PCR products showed correct size of each allele as expected. Conclusion Puml and Pum2 dCKO mice are established successfully, providing an useful conditional knockout animal model to study roles of PUF family at different stages and different cell types for spermatogenesis as well as for any other physiological process.