A preliminary study of conditional limb bud mesenchymal stem cell FKBP38 knockout mice
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摘要:
目的 构建并鉴定条件性肢芽间充质干细胞FKBP38基因敲除小鼠,为研究FKBP38基因在骨发育及骨关节炎中的作用机制提供动物模型。 方法 将Prrx1-Cre小鼠与FKBP38flox/flox交配繁殖出F1代小鼠,得到基因型为Prrx1-Cre(Fkbp38flox/+)小鼠;选基因型Prrx1-Cre(Fkbp38flox/+)的小鼠与FKBP38flox/flox小鼠杂交获得F2代小鼠;基因型为Prrx1-Cre(Fkbp38flox/flox)的小鼠即为本实验所需要构建模型小鼠。3周龄鉴定小鼠基因型,4周龄时用Western blot验证FKBP38基因敲除效果。 结果 F2代小鼠中包含基因敲除鼠,PCR结果显示基因型为Cre/FKBP38flox/flox,Western blot结果显示FKBP38基因敲除效果明显。 结论 基于Cre/loxp重组酶系统,繁殖出肢芽间充质干细胞FKBP38基因敲除鼠,为在动物水平上探究FKBP38在骨发育及骨关节炎的作用机制提供模型基础。 -
关键词:
- 肢芽间充质干细胞 /
- FKBP38 /
- Cre/loxp重组酶系统
Abstract:Objective To construct and identify conditional limb bud mesenchymal stem cell FKBP38 knockout mice, and provide an animal model for the mechanism of FKBP38 gene in bone development and osteoarthritis. Methods Prrx1-Cre mice were mated with FKBP38flox/flox to produce F1 mice. The genotype was Prrx1-Cre(Fkbp38flox/+) mice. The mice with genotype Prrx1-Cre(Fkbp38flox/+) were selected. FKBP38flox/flox mice were crossed to obtain F2 mice. Mice with genotype Prrx1-Cre(Fkbp38flox/flox) were used to construct model mice for the experiment. The mouse genotype was identified at 3 weeks of age. The knockdown effect of FKBP38 gene was verified by Western blot at 4 weeks of age. Results F2 mice contained knockout mice. The PCR results showed that the genotype was Cre/FKBP38flox/flox. The results of Western blot showed that the FKBP38 gene knockout effect was obvious. Conclusion Based on the Cre/loxp system, the limb bud mesenchymal stem cell FKBP38 knockout mouse is propagated to provide a model basis for exploring the mechanism of FKBP38 in bone development and osteoarthritis at the animal level. -
Key words:
- limb bud mesenchymal stem cells /
- FKBP38 /
- Cre/loxp recombinase system
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表 1 基因鉴定引物
Gene Primer (5′→3′) Length of product (bp) Prx1 F: TCCAATTTACTGACCGTACACCAA
R: CCTGATCCTGGCAATTCGGCTA500 FKBP38 F: TCATGCAGGCAGAGTGCACATAG
R: CTGGAACTGGAGACTGGCTCCAMT: 272WT: 217 -
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