Two-Hybrid Systems

1. Front Matter

   Pages i-xii

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2. Protein Interactomics by Two-Hybrid Methods

   • Soon Gang Choi, Aaron Richardson, Luke Lambourne, David E. Hill, Marc Vidal

   Pages 1-14

3. Yeast Hybrid Systems to Study Protein, Nucleic Acid, and Ligand Interactions

   1. Front Matter

      Pages 15-15

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   2. Making the Right Choice: Critical Parameters of the Y2H Systems

      • Jitender Mehla, J. Harry Caufield, Peter Uetz

      Pages 17-28

   3. Identification of Protein–Protein Interactions Using Pool-Array-Based Yeast Two-Hybrid Screening

      • Thomas Lathouwers, Jeroen Wagemans, Rob Lavigne

      Pages 29-48

   4. Detecting Interactions of Membrane Proteins: The Split-Ubiquitin System

      • Lisa Yasmin Asseck, Christopher Grefen

      Pages 49-60

   5. The Ras Recruitment System (RRS) for the Identification and Characterization of Protein–Protein Interactions

      • Ami Aronheim

      Pages 61-73

   6. A Bacterial Adenylate Cyclase-Based Two-Hybrid System Compatible with Gateway® Cloning

      • Macy G. Olson, Megan Goldammer, Emilie Gauliard, Daniel Ladant, Scot P. Ouellette

      Pages 75-96

   7. Analyses of Protein Interaction Networks Using Computational Tools

      • Shaowei Dong, Nicholas J. Provart

      Pages 97-117

   8. Options and Considerations When Using a Yeast One-Hybrid System

      • Jared A. Sewell, Juan I. Fuxman Bass

      Pages 119-130

   9. Screening Arrayed Libraries with DNA and Protein Baits to Identify Interacting Proteins

      • Rocío Sánchez-Montesino, Luis Oñate-Sánchez

      Pages 131-149

   10. Construction of Arabidopsis Transcription Factor ORFeome Collections and Identification of Protein–DNA Interactions by High-Throughput Yeast One-Hybrid Screens

       • S. Earl Kang, Ghislain Breton, Jose L. Pruneda-Paz

       Pages 151-182

   11. Transcription Factor-Centered Yeast One-Hybrid Assay

       • Xiaoyu Ji, Liuqiang Wang, Dandan Zang, Yucheng Wang

       Pages 183-194

   12. The Yeast Three-Hybrid System for Protein Interactions

       • Franziska Glass, Mizuki Takenaka

       Pages 195-205

   13. The Yeast Three-Hybrid System for Screening RNA-Binding Proteins in Plants

       • Sung Ki Cho, David J. Hannapel

       Pages 207-224

   14. Using Yeast Hybrid System to Identify Proteins Binding to Small Molecules

       • You Wang, David S. Letham, Peter C. L. John, Ren Zhang

       Pages 225-234

   15. Shuttling of Entire Libraries from an Entry Vector to a Destination Vector of the Gateway System

       • Alexander Heyl

       Pages 235-242

4. Experimental Approaches to Study Protein-Protein and Protein-DNA Interactions in Plants and Human Cells

   1. Front Matter

      Pages 243-243

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   2. Bimolecular Fluorescence Complementation with Improved Gateway-Compatible Vectors to Visualize Protein–Protein Interactions in Plant Cells

      • Shino Goto-Yamada, Kazumi Hikino, Mikio Nishimura, Tsuyoshi Nakagawa, Shoji Mano

      Pages 245-258

   3. Bimolecular Fluorescence Complementation to Visualize Protein–Protein Interactions in Human Cells Based on Gateway Cloning Technology

      • Adriana Lepur, Oliver Vugrek

      Pages 259-267

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.

• Y2H system
• Protein-protein interactions
• Gene expression
• Yeast biology
• Protein-DNA interaction
• Plant and mammalian cells

• Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM)–Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, Madrid, Spain

  Luis Oñate-Sánchez

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