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通过在植物中针对特定位置的双硫酸盐测序进行单基分辨率DNA甲基化分析

Kirti Pandey^1,2, Gaurav Zinta^3,4

¹Integrative Plant AdaptOmics Lab (iPAL), Biotechnology Division, CSIR-Institute of Himalayan Bioresource Technology (IHBT), Palampur, Himachal Pradesh, India.

Methods in molecular biology (Clifton, N.J.)

|October 6, 2025

概括

这项研究详细介绍了一种二硫酸测序协议,用于精确的,特定于植物的DNA甲基化分析. 这种方法为研究基因调节和对压力的表观遗传反应提供了单基底分辨率.

科学领域:

植物分子生物学植物分子生物学
表观遗传学在表观遗传学中,表观遗传学是指表观遗传学.
基因组学就是基因组学.

背景情况:

基因甲基化 (5mC) 是植物的一个关键的表观遗传标记,它调节基因表达,可转移元素和跨CG,CHG和CHH环境的应激反应.
对于特定位置的DNA甲基化分析,存在各种方法,每种方法都有不同的优缺点.

研究的目的:

介绍一个详细的,高精度的协议,用于特定位置的植物DNA甲基化分析.
为了突出二硫酸盐测序作为其单基分辨率和简单性的黄金标准方法.

主要方法:

使用EZ DNA甲基化-GoldTM套件进行DNA的选择性双硫酸盐转化,将非甲基化细胞因子转化为 uracil.
采用精心设计的原始剂 (例如,通过MethPrimer) 来对二硫酸盐转化DNA进行特定放大.
涉及PCR放大,克隆成二进制矢量,以及用于单基分辨率分析的桑格测序.

主要成果:

该协议能够精确地绘制特定位置的DNA甲基化模式,包括促进剂,增强剂和可转移元素.
实现单基分辨率,提供对表观遗传修饰的详细见解.

结论:

二硫酸盐测序为植物的基位特异性DNA甲基化分析提供了强大而准确的方法.
这种技术对于研究基因调节,表观遗传和作物改进策略非常有价值.

关键词:

双硫酸盐序列测序双硫酸盐序列测序通过DNA甲基化.表观遗传修饰是一种表观遗传修饰.基因调节基因调节甲基化分析的分析方法

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