DNA repair: a simple enzymatic assay for human cells
DNA repair: a simple enzymatic assay for human cells
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DNA 修复：一种简单的人体细胞酶测定法
DOI：
10.1080/09553007314551531
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发表时间：
1973
影响因子：
2.6
通讯作者：
R. Wilkins
中科院分区：
文献类型：
--
作者：
R. Wilkins
基于藤黄微球菌提取物中某些核酸内切酶在 DNA 损伤位点特异性反应并产生单链断裂的能力描述了一种测定方法。通过碱沉淀 DNA 来检测断裂。如果在细胞处理后孵育期间核酸内切酶敏感位点的数量减少，则假定发生 DNA 修复。据称，通过将藤黄杆菌提取物添加到部分破碎的细胞中，而不是分离纯化的 DNA，对现有技术进行了改进，使得样品可以在 30 分钟内处理，大部分操作都在 0 ℃ 下进行。每 1 亿道尔顿 DNA 中可以检测到的核酸内切酶敏感位点少至一到两个。从紫外线照射的人二倍体成纤维细胞获得的结果证明了该方法的潜力。 （英国）
An assay is described based on the ability of certain endonucleases in Micrococcus luteus extracts to react specifically at sites of DNA damage, and produce single-strand breaks. The breaks were detected by sedimenting the DNA through alkali. DNA repair was assumed to occur if the number of endonuclease sensitive sites decreased during post-treatment incubation of cells. It is claimed that improvements have been obtained in an established technique by adding M. luteus extract to partially disintegrated cells, instead of to isolated purified DNA, enabling samples to be processed in 30 min with most of the manipulations at 0 deg C. As few as one or two endonuclease-sensitive sites per 100 million daltons of DNA could be detected. The potential of the method is demonstrated with results obtained from uv-irradiated human diploid fibroblasts. (UK)