学术
小鼠Pkhd1条件性基因打靶载体的构建
杨季云[1] 周钦[2] 张铮[2] 郭红[2] 张思仲[1]
[1]四川大学华西医院医学遗传研究室,四川成都610041 [2]四川大学生物治疗国家重点实验室基因工程小鼠中心,四川成都610041
摘 要:
目的构建小鼠Pkhdl条件性基因打靶栽俸。为建立Pkhdl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号外正于的Pkhdl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Pkhdl第6号外正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Pkhdl基因条件性打靶载体符合设计要求。结论成功构建了小鼠Pkhdl条件性基因打靶载体，为建立Pkhdl基因条件性敲除小鼠打下了基础。[著者文摘]
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文章出处:
《四川医学》-2007年28卷1期 -1-3页
栏目信息:
文献标识码:
A
文章编号:
1004-0501（2007）01-0001-03
Construction of targeting vector for conditional knockout of murine Pkhdl
Yang Jiyun , Zhou Qin , Zhang Zheng , et al （West China Hospital, Sichuan University, Chengdu , Sichuan 610041, China）
Abstract:
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.[著者文摘]
Key words:
APPKD; PKHD1 ; conditional knockout; homologous recombination
基金资助:
国家重点基础研究发展规划项目（973计划）（CB522506）
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