Richardson Rebecca M, Pascal Steven M
Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, Virginia, USA.
J Bacteriol. 2025 Aug 21;207(8):e0012925. doi: 10.1128/jb.00129-25. Epub 2025 Aug 1.
Bacterial two-hybrid (B2H) systems offer a versatile platform for detecting protein-protein interactions (PPIs) . Originally developed to study bacterial transcriptional regulators, these systems have evolved to support a wide range of applications, including interaction mapping, domain analysis, and high-throughput screening. This review surveys the development and expansion of B2H systems across diverse biological contexts. We highlight technical considerations, platform-specific innovations, and recent adaptations that extend the utility of B2H systems in both prokaryotic and eukaryotic protein interaction research environments. By comparing different B2H variants and discussing their integration with complementary approaches, we provide a framework for leveraging these systems in modern PPI discovery.
细菌双杂交（B2H）系统为检测蛋白质-蛋白质相互作用（PPI）提供了一个多功能平台。这些系统最初是为研究细菌转录调节因子而开发的，如今已发展到支持广泛的应用，包括相互作用图谱绘制、结构域分析和高通量筛选。本文综述了B2H系统在不同生物学背景下的发展和扩展。我们重点介绍了技术考量、特定平台的创新以及最近的改进，这些改进扩展了B2H系统在原核和真核蛋白质相互作用研究环境中的实用性。通过比较不同的B2H变体并讨论它们与互补方法的整合，我们提供了一个在现代PPI发现中利用这些系统的框架。
