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Bacterial two-hybrid systems evolved: innovations for protein-protein interaction research
Rebecca M Richardson1, Steven M Pascal1
1Department of Chemistry and Biochemistry, Old Dominion University, Norfolk, Virginia, USA.
Abstract:
Bacterial two-hybrid (B2H) systems offer a versatile platform for detecting protein-protein interactions (PPIs) in vivo. 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.
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