使用胞嘧啶延伸分析法分析全球基因组甲基化

Analysis of Global Genome Methylation Using the Cytosine-Extension Assay.

作者信息

Bilichak Andriy, Kovalchuk Igor

机构信息

Agriculture and Agri-Food Canada, Lethbridge Research Centre, Lethbridge, AB, Canada.

Department of Biological Sciences, University of Lethbridge, 4401 University Drive, Lethbridge, AB, Canada, T1K 3M4.

出版信息

Methods Mol Biol. 2017;1456:73-79. doi: 10.1007/978-1-4899-7708-3_6.

DOI:10.1007/978-1-4899-7708-3_6

PMID:27770358

Abstract

DNA methylation is a reversible covalent chemical modification of DNA intended to regulate chromatin structure and gene expression in a cell- and tissue-specific manner and in response to the environment. Cytosine methylation is predominantly occurring in plants, and cytosine nucleotides in plants can be methylated at symmetrical (CpG and CpHpG) and nonsymmetrical sites. Although there exists a number of various methods for the detection of cytosine methylation, most of them are either laborious or expensive or both. Here, we describe a quick inexpensive method for the analysis of global genome methylation using a cytosine-extension assay. The assay can be used for the analysis of the total level of CpG, CpHpG, and CpHpH methylation in a given sample of plant DNA.

摘要

DNA甲基化是一种可逆的DNA共价化学修饰，旨在以细胞和组织特异性方式并响应环境来调节染色质结构和基因表达。胞嘧啶甲基化主要发生在植物中，植物中的胞嘧啶核苷酸可在对称位点（CpG和CpHpG）和非对称位点发生甲基化。尽管存在多种检测胞嘧啶甲基化的方法，但其中大多数方法要么费力，要么昂贵，要么两者兼具。在此，我们描述了一种使用胞嘧啶延伸测定法快速且廉价地分析全基因组甲基化的方法。该测定法可用于分析给定植物DNA样品中CpG、CpHpG和CpHpH甲基化的总水平。