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This detailed book explores the numerous applications of the yeast two-hybrid (Y2H) system, one of the most commonly used methods to detect protein-protein interactions. Beginning with an overview, the volume then continues by elucidating different methodologies to detect protein-protein interactions in yeast nucleus, membrane, cytoplasm, and bacteria, computational analyses of protein interaction networks, technical variations in yeast one-hybrid (Y1H) systems together with classical and more elaborated yeast methods to detect protein-DNA interactions, as well as protocols to analyze ternary protein interactions and RNA-protein and ligand-protein interactions. The book concludes with a section on methods in plant and mammalian cells that can be used to identify novel protein-protein and protein-DNA interactions and test (qualitatively and/or quantitatively) those observed in yeast. Written for the highly successful Methods in Molecular Biology series, chapters include thekind of detail and key implementation advice that leads to excellent results in the lab.
Authoritative and practical, Two-Hybrid Systems: Methods and Protocols serves as an ideal guide to the uses of this exceedingly valuable technique.
List of contents
Protein Interactomics by Two-Hybrid Methods.- Making the Right Choice: Critical Parameters of the Y2H Systems.- Identification of Protein-Protein Interactions Using Pool-Array-Based Yeast Two-Hybrid Screening.- Detecting Interactions of Membrane Proteins: The Split-Ubiquitin System.- The Ras Recruitment System (RRS) for the Identification and Characterization of Protein-Protein Interactions.- A Bacterial Adenylate Cyclase-Based Two-Hybrid System Compatible with Gateway® Cloning.- Analyses of Protein Interaction Networks Using Computational Tools.- Options and Considerations When Using a Yeast One-Hybrid System.- Screening Arrayed Libraries with DNA and Protein Baits to Identify Interacting Proteins.- Construction of Arabidopsis Transcription Factor ORFeome Collections and Identification of Protein-DNA Interactions by High-Throughput Yeast One-Hybrid Screens.- Transcription Factor-Centered Yeast One-Hybrid Assay.- The Yeast Three-Hybrid System for Protein Interactions.- The Yeast Three-Hybrid System for Screening RNA-Binding Proteins in Plants.- Using Yeast Hybrid System to Identify Proteins Binding to Small Molecules.- Shuttling of Entire Libraries from an Entry Vector to a Destination Vector of the Gateway System.- Bimolecular Fluorescence Complementation with Improved Gateway-Compatible Vectors to Visualize Protein-Protein Interactions in Plant Cells.- Bimolecular Fluorescence Complementation to Visualize Protein-Protein Interactions in Human Cells Based on Gateway Cloning Technology.- KISS: A Mammalian Two-Hybrid Method for In Situ Analysis of Protein-Protein Interactions.- Coimmunoprecipitation of Interacting Proteins in Plants.- Analysis of Protein-Protein Interaction by Co-IP in Human Cells.- Tandem Affinity Purification of Protein Complexes from Arabidopsis Cell Cultures.- Transient Transactivation Studies in Nicotiana benthamiana Leaves.- The Use of the Chromatin Immunoprecipitation Technique for In Vivo Identification of Plant Protein-DNA Interactions.- Chromatin Immunoprecipitation for Identification of Protein-DNA Interactions in Human Cells.
Summary
This detailed book explores the numerous applications of the yeast two-hybrid (Y2H) system, one of the most commonly used methods to detect protein-protein interactions. Beginning with an overview, the volume then continues by elucidating different methodologies to detect protein-protein interactions in yeast nucleus, membrane, cytoplasm, and bacteria, computational analyses of protein interaction networks, technical variations in yeast one-hybrid (Y1H) systems together with classical and more elaborated yeast methods to detect protein-DNA interactions, as well as protocols to analyze ternary protein interactions and RNA-protein and ligand-protein interactions. The book concludes with a section on methods in plant and mammalian cells that can be used to identify novel protein-protein and protein-DNA interactions and test (qualitatively and/or quantitatively) those observed in yeast. Written for the highly successful Methods in Molecular Biology series, chapters include thekind of detail and key implementation advice that leads to excellent results in the lab.
Authoritative and practical, Two-Hybrid Systems: Methods and Protocols serves as an ideal guide to the uses of this exceedingly valuable technique.
