摘要
目的构建小鼠Pkhd1条件性基因打靶载体,为建立Pkhd1条件性敲除小鼠模型奠定基础。方法以正常小鼠(129xl/SvJ)基因组DNA为模板,扩增小鼠包括第6号外显子的Pkhd1基因部分序列,通过引入LoxP和Neo基因等步骤,建立条件性敲除Pkhd1第6号外显子的条件性基因打靶载体。结果经多个限制性核酸内切酶酶切鉴定和测序证实,构建的小鼠Pkhd1基因条件性打靶载体符合设计要求。结论成功构建了小鼠Pkhd1条件性基因打靶载体,为建立Pkhd1基因条件性敲除小鼠打下了基础。
Objective To construct the targeting vector for conditional knockout for polycystic kidney and hepatic disease 1 gene （Pkhdl） .Methods A 9.3 kb Pkhdl genomie DNA frasment containing exon 6, obtained from routine genomie DNA（ 129xl/SvJ ）by PCR, was inserted into T vector（ pCB2. 1-TOPO）whieh was used as the homologous ann. Two LoxP was inserted into intron 5 and 6 for conditional knockout of exon 6 of Pkhdl. Resnlts The correct structure of the targeting vector was confu-mcd by restriction enzyme digestion and sequencing analysis. Conclusion A targeting vector for conditional knockout of routine Pkhdl has been successfully constructed.The construction of targeting vector paved the way for conditional knockout mouse strain generated by targeted mutation of Pkhdl.
出处
《四川医学》
CAS
2007年第1期1-3,共3页
Sichuan Medical Journal
基金
国家重点基础研究发展规划项目(973计划)(CB522506)
