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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 identify a mouse model of conditional knockdown of Pumilio1 / Pumilio2 (Pum1 and Pum2) genes and lay a foundation for further study on the biological function of PUF family in mammalian spermatogenesis. Methods Two strains of mice with conditional knockout of Pum1 and conditional knockout of Pum2 were crossed and propagated. The two breeds Pum1 and Pum2 were successfully homozygous. The offspring were numbered by pruning, genomic DNA was extracted from toe or rat tail of offspring mice, genotypes were identified by PCR and agarose gel electrophoresis. Results The mouse model of conditional knockout of Pum1 and Pum2 was successfully propagated, and more mice with conditional knockdown of the dual gene of PUF family were obtained. Conclusion The mouse model of conditional knockout of Pum1 and Pum2 genes was established successfully. The correct strategy of mating and breeding and identification of the two genes were effective ways to obtain double gene knockout mice with PUF family.