上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

Tex101条件性基因敲除小鼠模型的建立和表型鉴定

冯贵莲，舒 洋，王保曼，朱于非，胡兰靛   

  1. 上海交通大学 医学院/中科院上海生命科学研究院 健康科学研究所分子遗传学研究组， 上海 200025
  • 出版日期:2014-01-28 发布日期:2014-01-29
  • 作者简介:冯贵莲（1986—）， 女， 硕士; 电子信箱: fengguilian@163.com。
  • 基金资助:

    国家重点基础研究发展计划（“973”计划）（2011CB510102）

Generation of Tex101 conditional knockout mice and phenotype identification

FENG Gui-lian, SHU Yang, WANG Bao-man, ZHU Yu-fei, HU Lan-dian   

  1. Laboratory of Molecular Genetics, Institute of Health Sciences, Shanghai Jiao Tong University School of Medicine/Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025,  China
  • Online:2014-01-28 Published:2014-01-29
  • Supported by:

    National Basic Research Program of China, 973 Program, 2011CB510102

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摘要：

目的 建立Tex101（Testis expressed gene 101)条件性基因敲除小鼠模型并进行表型鉴定。方法 构建针对小鼠Tex101基因2、3号外显子的重组载体，通过电穿孔的方法转染胚胎干细胞(ES细胞)，用正负筛选法筛选阳性ES细胞并进行PCR鉴定。利用显微注射把发生正确同源重组的ES细胞注射进C57BL/6J小鼠囊胚，移入受体小鼠子宫。将得到的嵌合体小鼠与C57BL/6J雌鼠交配获得Tex101-LoxP转基因小鼠。该小鼠与EⅡa-Cre 转基因小鼠杂交，通过子代自交获得Tex101条件性基因敲除（cKO)小鼠并进行初步分析。结果 得到2只Tex101 cKO雄鼠。冰冻切片观察发现Tex101 cKO雄鼠睾丸形态和大小与野生型小鼠相比无明显差异。结论 成功建立了Tex101条件性基因敲除小鼠模型，并初步验证了该基因对雄性小鼠睾丸大小和形态无明显影响，为进一步研究Tex101基因功能奠定了基础。

关键词: Tex101, 条件性基因敲除, 胚胎干细胞, EⅡa-Cre转基因小鼠

Abstract:

Objective To generate testis expressed gene 101 (Tex101) conditional knockout mice model, and to identify the phenotype. Methods The conditional knockout vector targeting exon 2 and exon 3 of Tex101 were generated by molecular cloning, and then the electroporation of embryonic stem (ES) cells was carried out. The targeted ES cells were screened by positive-negative selection and PCR, and the correct targeted ES cells were microinjected into blastula of C57BL/6J mice. Chimerical mice were generated after transplanted the blastula into the host mice, and then the Tex101-LoxP transgenic mice were generated. Tex101 cKO mice could be obtained by crossing between EⅡa-Cre mice and Tex101-LoxP transgenic mice. The genotype was identified by PCR and phenotype by frozen section. Results We got 2 male Tex101 cKO mice and the frozen section showed no discrepancy of testis morphology between WT and cKO mice. Also, there was no obvious difference of the testis weight. Conclusion The Tex101 conditional knockout mice were successfully obtained, which showed no obvious influence on the testis. This animal model provided the foundation for further study of Tex101 in vivo.

Key words: Tex101, conditional knockout, embryonic stem cell, EⅡa-Cre transgenic mouse