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用于基因组稳定性研究的基于报告基因的重组系。

Reporter gene-based recombination lines for studies of genome stability.

作者信息

Kathiria Palak, Kovalchuk Igor

机构信息

Department of Biological Sciences, University of Lethbridge, Lethbridge, AB, Canada.

出版信息

Methods Mol Biol. 2010;631:243-52. doi: 10.1007/978-1-60761-646-7_18.

Abstract

Homologous recombination is a double-strand break repair mechanism operating in somatic cells and involved in meiotic crossovers in plants. It is responsible for the maintenance of genome stability and thus plays a crucial role in adaptation to stress. Recombination between homologous loci is believed to be regulated in part by epigenetic machinery such as methylation. Therefore, the recombination frequency at a specific locus can reflect the chromatin status.Several reporter gene-based recombination constructs have been developed to study HR frequencies in plants. Among them, the luciferase and beta-glucuronidase-based recombination reporter systems are the most widely used. Here, we explain how reporter gene recombination assays operate and in which applications they are used. We also present a conceptually new system for analysis of sequence-specific recombination frequency. These assays can be effectively used for analysis of locus-specific endogenous and stress-induced recombination frequencies.

摘要

同源重组是一种在体细胞中起作用的双链断裂修复机制,参与植物减数分裂中的交叉互换。它负责维持基因组稳定性,因此在适应胁迫方面起着关键作用。同源位点之间的重组被认为部分受表观遗传机制(如甲基化)调控。因此,特定位点的重组频率可以反映染色质状态。为研究植物中的同源重组频率,已开发出几种基于报告基因的重组构建体。其中,基于荧光素酶和β-葡萄糖醛酸酶的重组报告系统使用最为广泛。在此,我们解释报告基因重组测定如何运作以及它们用于哪些应用。我们还提出了一种概念上全新的分析序列特异性重组频率的系统。这些测定可有效用于分析位点特异性内源性和胁迫诱导的重组频率。

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