优化的DNA亲和力净化测序确定了转录因子的相对结合亲和力
Katharina Schiller1,2, Anja Meierhenrich1,2, Sanja Zenker1,2
1Computational Biology, Faculty of Biology, Bielefeld University, 33615 Bielefeld, Germany.
Biology methods & protocols
|December 8, 2025
概括
DNA亲和净化测序 (DAP-seq) 是一种可扩展的方法来分析与DNA结合的转录因子. 这项研究优化了DAP-seq协议,并为分析任何物种的蛋白质-DNA相互作用提供了指南.
科学领域:
- 分子生物学分子生物学
- 基因组学就是基因组学.
- 生物信息学是一种生物信息学.
背景情况:
- 转录因子 (TFs) 通过与特定的DNA序列结合来调节基因表达.
- 评估TF-DNA结合亲和力对于理解基因调节至关重要.
- 现有的方法可能缺乏可扩展性或需要进行广泛的优化.
研究的目的:
- 为量化TF-DNA相互作用提供一个优化的DNA亲和力净化测序 (DAP-seq) 协议.
- 为DAP-seq数据的生物信息分析提供实用指南.
- 提高TF结合亲缘关系研究的可复制性和可扩展性.
主要方法:
- 在体外表达标记的转录因子 (例如,TF::HaloTag).
- 基因组DNA的碎片化和适配器绑定.
- 使用亲和标签和随后的测序来净化TF结合的DNA.
- 量化步骤,使图书馆的大小均化,并提高可复制性.
主要成果:
- 一个优化的DAP-seq协议用于蛋白质-DNA相互作用的相对量化.
- 一个全面的生物信息管道用于识别TF绑定站点.
- 分析复制强度,控制特征和相对结合亲和力的方法.
结论:
- 优化的DAP-seq协议对于研究跨物种TF-DNA相互作用是高效和可扩展的.
- 提供的数据分析指南有助于在各种研究环境中应用DAP-seq.
- 这种方法通过能够对TF结合亲缘关系进行可靠的量化来促进对基因调节的理解.
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