title: A bidirectional fluorescent two-hybrid system for monitoring protein-protein interactions.
abstract: Two-hybrid systems have been the cornerstone of research into protein-protein interactions, but these systems typically rely on life/death reporters that put additional selective pressure on the host organism, and potentially lead to false positives. Here we report a bidirectional fluorescence-based bacterial two-hybrid system that enables both the association and dissociation of a given protein-protein interaction to be monitored. The functionality of this system and its compatibility with FACS screening are demonstrated in the forward and reverse direction using known interacting protein-partners and their cyclic peptide inhibitors. The reported fluorescent two-hybrid system may be used in the forward direction for the identification of interacting protein partners, or as a reverse two-hybrid system for the high-throughput identification of protein-protein interaction inhibitors.
authors.affiliation: Chemistry, University of Southampton, Southampton, SO17 1BJ, UK.
authors.forename: Ida Karin
authors.initials: IK
authors.lastname: Nordgren
authors.forename: Ali
authors.initials: A
authors.lastname: Tavassoli
mesh_terms: D055808:Drug Discovery; D005434:Flow Cytometry; D015870:Gene Expression; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D025941:Protein Interaction Mapping / Q000379:methods; D020798:Two-Hybrid System Techniques