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目的构建小鼠ADAM10条件性基因打靶载体,为建立ADAM10条件性敲除小鼠模型奠定基础。方法以正常小鼠(129/Svj)基因组DNA为模板,采用长片段PCR方法,扩增小鼠ADAM10基因第2外显子及其侧翼序列。通过引入LoxP位点和TK基因等步骤,获得ADAM10条件性基因打靶载体。结果经限制性内切酶酶切鉴定和测序证实,构建的小鼠ADAM10基因条件性打靶载体符合设计要求。结沦成功构建了小鼠ADAM10条件性基因打靶载体,为建立ADAM10条件性基因敲除小鼠打下了基础。
Objective To construct mouse ADAM10 conditional gene targeting vector and lay the foundation for the establishment of ADAM10 conditional knockout mouse model. Methods The genomic DNA of normal mouse (129 / Svj) was used as a template to amplify the exon 2 of ADAM10 gene and its flanking sequence using long fragment PCR. ADAM10 conditional gene targeting vector was obtained through the steps of introducing LoxP site and TK gene. Results Restriction endonuclease digestion and sequencing confirmed that the constructed mouse ADAM10 gene conditional targeting vector met the design requirements. Conclusions Successfully constructed mouse ADAM10 conditional gene targeting vector, laid the foundation for the establishment of ADAM10 conditional knockout mice.