Foerster Andrea M, Mittelsten Scheid Ortrun
Gregor Mendel Institute of Molecular Plant Biology, Austrian Academy of Sciences, Vienna, Austria.
Methods Mol Biol. 2010;631:1-11. doi: 10.1007/978-1-60761-646-7_1.
Methylation of cytosines is a very important epigenetic modification of genomic DNA in many different eukaryotes, and it is frequently involved in transcriptional regulation of genes. In plants, DNA methylation is regulated by a complex interplay between several methylating and demethylating enzymes. Analysis of the resulting cytosine methylation patterns with the highest resolution is achieved after sodium bisulfite treatment, deaminating nonmethylated cytosines to uracil. Subsequent PCR and sequence analysis of individual amplicons displays the degree, position, and sequence context of methylation of every cytosine residue in individual genomic sequences. We describe the application of bisulfite sequencing for the analysis of DNA methylation at defined individual sequences of plant genomic DNA.
胞嘧啶甲基化是许多不同真核生物基因组DNA非常重要的表观遗传修饰，并且它经常参与基因的转录调控。在植物中，DNA甲基化由几种甲基化和去甲基化酶之间复杂的相互作用调控。用亚硫酸氢钠处理后，将未甲基化的胞嘧啶脱氨基转化为尿嘧啶，从而实现对所得胞嘧啶甲基化模式的最高分辨率分析。随后对单个扩增子进行PCR和序列分析，可显示单个基因组序列中每个胞嘧啶残基的甲基化程度、位置和序列背景。我们描述了亚硫酸氢盐测序在分析植物基因组DNA特定单个序列处DNA甲基化中的应用。
