GPR126条件性基因敲除载体的快速构建

中国生物工程杂志

2013, Vol. 33

Issue (4): 106-113

技术与方法

GPR126条件性基因敲除载体的快速构建

叶湘漓^1,2, 李大力²

1. 湖南师范大学医学院长沙 410013;
2. 华东师范大学生命科学学院生命医学研究所上海 200241

Rapid Construction of GPR126 Conditional Gene-targeting Vector

YE Xiang-li^1,2, LI Da-li²

1. College of Medicine, Hunan Normal University, Changsha 410013, China;
2. Institute of Biomedical Sciences, East China Normal University, Shanghai 200241, China

全文: PDF(625 KB) HTML

摘要： 基因敲除小鼠模型是在动物体内研究基因功能的重要和可靠的手段之一,而构建用于同源重组的基因敲除载体是构建基因敲除小鼠模型的关键一步。条件性基因敲除技术将针对靶基因的修饰作用限制在小鼠的特定发育时期或特定类型的组织细胞中,通过加入具有位点特异性的重组系统,该修饰作用在时间和空间上均处于可调控的状态;条件性基因敲除技术既可以避免某些具有重要功能基因的敲除所带来的早期胚胎致死问题,还可用于精确分析某些多效基因在不同组织或不同发育阶段的特定功能差异。通过合理利用改良的Red重组系统,将两个LoxP位点快速准确地插入到了目标位置,实现了GPR126条件性基因敲除载体的快速构建,为后续的构建GPR126基因敲除小鼠模型、在动物体内研究基因的功能奠定了基础。

关键词： 条件性基因敲除; 打靶载体; Red重组酶

Abstract: Generation of specific gene knockout mouse model is a reliable technique for studying the role of genes in mammalian model organism. One of the key steps to acquire a gene knockout mouse is to construct a targeting vector for homologous recombination in mouse embryonic stem cells. For some genes, the mutants will die in uteri owing to their critical roles in embryonic development, or the mutant mice may have different phenotypes according to the period of development and types of tissues. It is difficult to study these genes by the conventional knockout approaches. Thereby the conditional knockout strategy had been developed for its advantages to circumvent the embryonic lethality problem and to investigate gene function temporally and spatially. Using modified Red homologous recombineering system, the two LoxP sites were inserted into the target position accurately and the GPR126 conditional gene-targeting vector was rapidly constructed. The successful generation of GPR126 conditional knockout construct will be helpful for the subsequent production of knockout mouse model and exploration of the function of GPR126 in mice.

Key words: Conditional gene knockout Gene targeting vector Red recombinase

收稿日期: 2012-07-04 出版日期: 2013-04-25

ZTFLH:

Q789

基金资助: 国家自然科学基金(31171318,81270291);湖南省自然科学基金(12JJ3117);湖南省高等学校科学研究(12C0200)资助项目

通讯作者: 李大力 E-mail: dlli@bio.ecnu.edu.cn

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引用本文:

叶湘漓, 李大力. GPR126条件性基因敲除载体的快速构建[J]. 中国生物工程杂志, 2013, 33(4): 106-113.

YE Xiang-li, LI Da-li. Rapid Construction of GPR126 Conditional Gene-targeting Vector. China Biotechnology, 2013, 33(4): 106-113.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I4/106

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