S100a4-Cre~× Ddr2^(flox/flox)条件敲除小鼠的繁育及基因型鉴定

Reproduction and Genotyping of the S100a4-Cre~× Ddr2^(flox/flox) Conditional Knock-Out Mice

目的:利用Cre-LoxP重组酶系统构建成纤维细胞中Ddr2特异性敲除的纯合子小鼠并鉴定,为进一步研究Ddr2在肺纤维化中的作用提供基础。方法:将购买的Ddr2 loxp小鼠和成纤维细胞特异表达的S100a4-Cre小鼠分别繁殖与鉴定,然后将两种小鼠杂交与鉴定,最终得到基因型为Cre~× Ddr2^(flox/flox)的纯合子小鼠就是在成纤维细胞中Ddr2条件性敲除小鼠。结果:成功繁殖并用PCR技术准确鉴定了基因型为Cre~× Ddr2^(flox/flox)的纯合子小鼠,Western blot结果表明纯合子小鼠的肺成纤维细胞中Ddr2已缺失。结论:本研究利用Cre-LoxP系统成功构建了在成纤维细胞中Ddr2特异性敲除的纯合子小鼠,为进一步研究Ddr2在肺纤维化发展中的机制提供了研究平台。

Objective： To generate and identify the fibroblast specific Ddr2 knockout homozygous mice, for the research of the role of the Ddr2 gene on the pulmonary fibrosis. Methods： First, The introduced Ddr2 loxp mice and the S100 a4-Cre mice which expressed fibroblast cell-specific had reproduced and genotyped respectively, then the two kinds of mice were crossed, finally the off-spring mice which genotyping CreDdr2^ flox/floxwere the homozygous mice. Results： CreuDdr2^flox/floxmicewere breed successfully and their genotype were identified accurately by PCR. Conclusions： We successfully generated fibroblast specific Ddr2 gene knockout homozygous mice through Cre-LoxPtechnology, that can supply the tool for research the role of the Ddr2 gene on pulmonary fibrosis.