A novel combined bisulfite UDG assay for selective 5-methylcytosine detection
A novel combined bisulfite UDG assay for selective 5-methylcytosine detection
复制标题
一种用于选择性 5-甲基胞嘧啶检测的新型组合亚硫酸氢盐 UDG 测定法
DOI：
10.1016/j.talanta.2013.09.026
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发表时间：
2013-12-15
期刊：
影响因子：
6.1
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
--
作者：
Huang, Rong;Wang, Jiaqi;Zhou, Xiang
DNA修饰是一种重要的表观遗传事件，它在很大程度上影响着基因与转录因子和其他DNA结合蛋白的结合。在DNA修饰中，甲基化，尤其是胞嘧啶甲基化由于其导致肿瘤抑制基因表达的沉默而引起广泛的研究。在这项工作中，一种新的组合亚硫酸氢盐尿嘧啶-DNA糖基化酶（UDG）测定已开发的基础上，亚硫酸氢盐修饰，以产生尿嘧啶从胞嘧啶，随后UDG介导的尿嘧啶消除和最终的DNA切割在碱性条件下。该策略可用于选择性地检测5-甲基胞嘧啶残基的确切数目和位点，而不管序列背景如何。此外，它提供了DNA甲基化水平在很宽范围内的线性定量结果。(C)2013 Elsevier B. V.保留所有权利。
DNA modification, a significant epigenetic event, largely affects genes' binding with the transcription factors and some other DNA binding proteins. Among DNA modifications, methylation, especially cytosine methylation is of great importance and attracts extensive studies as it leads to the silence of tumor-suppressor gene expression. In this work, a novel combined bisulfite Uracil-DNA glycosylase (UDG) assay has been developed on the basis of bisulfite modification to generate uracil from cytosine, subsequent UDG-mediated uracil elimination and ultimate DNA cleavage in alkaline condition. This strategy can be used to selectively detect exact number and loci of 5-methylcytosine residues regardless of sequence context. Moreover, it provides linear quantitative results of DNA methylation level across a wide range. (C) 2013 Elsevier B.V. All rights reserved.