Paiano Aurora, Margiotta Azzurra, De Luca Maria, Bucci Cecilia
Department of Biological and Environmental Sciences and Technologies (DiSTeBA), University of Salento, Lecce, Italy.
Present address: Department of Biosciences, Centre for Immune Regulation, University of Oslo, Oslo, Norway.
Curr Protoc Protein Sci. 2019 Feb;95(1):e70. doi: 10.1002/cpps.70. Epub 2018 Aug 21.
This article describes the general method to perform the classical two-hybrid system. Although it has already been more than 25 years since this technique was developed, it still represents one of the best and most inexpensive, time saving, and straightforward methods to identify and study protein-protein interactions. Indeed, this system can be easily used to identify interacting proteins for a given protein, to check interactions between two known proteins, or to map interacting domains. Most of the interactions revealed using the two-hybrid assay have been proven to be binary direct interactions. Data comparison with other systems, such as mass spectrometry, have demonstrated that this system is at least as reliable. In fact, its use is increasing with time, and at present numerous variants of the yeast two-hybrid assay have been developed, including high-throughput systems that promote the generation of a proteome-scale map of protein-protein interactions in specific system. © 2018 by John Wiley & Sons, Inc.
本文描述了进行经典双杂交系统的一般方法。尽管这项技术自开发以来已有25年多的时间，但它仍然是鉴定和研究蛋白质-蛋白质相互作用的最佳、最经济、最省时且最直接的方法之一。实际上，该系统可轻松用于鉴定给定蛋白质的相互作用蛋白、检查两个已知蛋白质之间的相互作用或绘制相互作用结构域。使用双杂交分析揭示的大多数相互作用已被证明是二元直接相互作用。与其他系统（如质谱）的数据比较表明，该系统至少同样可靠。事实上，随着时间的推移，其应用越来越广泛，目前已经开发出许多酵母双杂交分析的变体，包括促进在特定系统中生成蛋白质-蛋白质相互作用的蛋白质组规模图谱的高通量系统。© 2018约翰威立国际出版公司。
