title: Using the yeast two-hybrid system to identify protein-protein interactions.
abstract: The yeast two-hybrid system is currently one of the most standardized protein interaction mapping techniques. The rationale of the yeast two-hybrid system relies on the physical separation of the DNA-binding domain from the transcriptional activation domain of several transcription factors. The protein of interest (bait) is fused to a DNA-binding domain, and complementary DNA (cDNA) library-encoded proteins are fused to a transcriptional activation domain. When a protein encoded by the cDNA library binds to the bait, both activities of the transcription factor are rejoined resulting in transcription from a reporter gene. Here, we describe protocols to test interactions between two individual proteins and to look for novel interacting partners by screening a single protein or domain against a library of other proteins using a GAL4 based yeast two-hybrid system.
authors.affiliation: Departamento de Biología Celular, Genética y Fisiología, Instituto de Hortofruticultura Subtropical y Mediterránea "La Mayora", Universidad de Málaga-Consejo Superior de Investigaciones Científicas (IHSM-UMA- CSIC), Universidad de Málaga, Málaga, Spain.
authors.forename: Edgar
authors.initials: E
authors.lastname: Rodríguez-Negrete
authors.forename: Eduardo R
authors.initials: ER
authors.lastname: Bejarano
authors.forename: Araceli G
authors.initials: AG
authors.lastname: Castillo
mesh_terms: D015723:Gene Library; D005822:Genetic Vectors; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000254:growth & development / Q000378:metabolism; D014170:Transformation, Genetic; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D001616:beta-Galactosidase