Fuxman Bass Juan I, Reece-Hoyes John S, Walhout Albertha J M
Program in Systems Biology, University of Massachusetts Medical School, Worcester, Massachusetts 01605.
Cold Spring Harb Protoc. 2016 Dec 1;2016(12):pdb.top077669. doi: 10.1101/pdb.top077669.
An important question when studying gene regulation is which transcription factors (TFs) interact with which cis-regulatory elements, such as promoters and enhancers. Addressing this issue in complex multicellular organisms is challenging as several hundreds of TFs and thousands of regulatory elements must be considered in the context of different tissues and physiological conditions. Yeast one-hybrid (Y1H) assays provide a powerful "gene-centered" method to identify the TFs that can bind a DNA sequence of interest. In this introduction, we describe the basic principles of the Y1H assay and its advantages and disadvantages and briefly discuss how it is complementary to "TF-centered" methods that identify protein-DNA interactions for a known protein of interest.
在研究基因调控时,一个重要的问题是哪些转录因子(TFs)与哪些顺式调控元件相互作用,例如启动子和增强子。在复杂的多细胞生物中解决这个问题具有挑战性,因为在不同组织和生理条件的背景下,必须考虑数百种转录因子和数千个调控元件。酵母单杂交(Y1H)试验提供了一种强大的“以基因为中心”的方法,用于鉴定能够结合感兴趣DNA序列的转录因子。在本引言中,我们描述了Y1H试验的基本原理及其优缺点,并简要讨论了它如何与“以转录因子为中心”的方法互补,后者用于鉴定针对已知感兴趣蛋白质的蛋白质-DNA相互作用。