癌变·畸变·突变 ›› 1999, Vol. 11 ›› Issue (4): 161-164.doi: 10.3969/j.issn.1004-616x.1999.04.002

• 论著 • 上一篇    下一篇

含小鼠糖皮质激素受体第二外显子基因片段的克隆和结构分析

李　坚 胡以平　傅继梁   

  1. 第二军医大学基础部生物教研室、医学分子遗传学开放实验室　上海　200433
  • 收稿日期:1999-04-26 修回日期:1999-05-05 出版日期:1999-07-30 发布日期:1999-07-30

CLONING AND STRUCTURE ANALYSIS OF GENE FRAGMENT BEARING MOUSE GLUCOCORTICOID RECEPTOR GENE EXON Ⅱ

Li Jian , Hu Yiping , Fu J iliang   

  1. Department of Biology and Open L ab of Medical Molecular Genetics , Second Military Medical University , S hanghai 　200433
  • Received:1999-04-26 Revised:1999-05-05 Online:1999-07-30 Published:1999-07-30

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摘要： 克隆适于构建可调控基因打靶载体的小鼠糖皮质激素受体( GR) 基因片段,为建立糖皮质激素受体基因缺陷型小鼠模型奠定基础。用PCR 扩增小鼠GR 基因第二外显子上562bp 的核酸片段作为探针,筛选小鼠基因组文库,共获得6 个阳性克隆。对C10 克隆进行详细的测序和酶切图谱和Southern 杂交分析后获得一6. 5kb 的基因片段。该片段含完整的GR 基因第二外显子,左右分别有319kb 和114kb 的DNA 片段可作为下一步构建基因打靶载体的同源臂。

关键词: 基因打靶, 糖皮质激素受体基因

Abstract: To clone a genomic DNA f ragment containing mouse glucocorticoid receptor ( GR) gene for the const ruction of an inducible gene targeting vector used in the establishment of a GR gene dificient mouse model. The mouse genomic library was screened by using a 562bp PCR product from GR gene exon Ⅱas a probe. Six positive clones were got after screening. The DNA of positive clones was profoundly characterized by sequencing , rest riction mapping and Southern blotting to get the right f ragment . A 6. 5 kb DNA f ragment was separated from the C10 clone. The gene f ragment contains entire GR gene exon Ⅱ, and two flanking f ragment s which are 3. 9 kb and 1. 4 kb respectively can be used as homologous arm in the const ruction of the targeting vector.

Key words: gene targeting, glucocorticoid receptor gene

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