通过连接介导的聚合酶链反应在DNA序列水平上测量紫外线损伤的形成和修复。
Measuring the formation and repair of UV damage at the DNA sequence level by ligation-mediated PCR.
作者信息
Besaratinia Ahmad, Pfeifer Gerd P
机构信息
Division of Biology, Beckman Research Institute of the City of Hope, Duarte, CA, USA.
出版信息
Methods Mol Biol. 2012;920:189-202. doi: 10.1007/978-1-61779-998-3_14.
Abstract
The formation and repair of DNA damage at specific locations in the genome is modulated by DNA sequence context, by DNA cytosine-5 methylation patterns, by the transcriptional status of the locus and by proteins associated with the DNA. The only method currently available to allow precise sequence mapping of DNA lesions in mammalian cells is the ligation-mediated polymerase chain reaction (LM-PCR) technique. We provide an update on technical details of LM-PCR. LM-PCR can be used, for example, for mapping of ultraviolet (UV) light-induced DNA photoproducts such as cyclobutane pyrimidine dimers.
摘要
基因组中特定位置的DNA损伤的形成和修复受到DNA序列背景、DNA胞嘧啶-5甲基化模式、基因座的转录状态以及与DNA相关的蛋白质的调节。目前唯一可用于对哺乳动物细胞中的DNA损伤进行精确序列定位的方法是连接介导的聚合酶链反应（LM-PCR）技术。我们提供了LM-PCR技术细节的最新情况。例如，LM-PCR可用于定位紫外线（UV）诱导的DNA光产物，如环丁烷嘧啶二聚体。
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