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Two-Hybrid Systems

Methods and Protocols

  • Book
  • © 2018

Overview

Editors:
  1. Luis Oñate-Sánchez

    1. 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

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  • Includes cutting-edge methods and protocols for the study of two-hybrid systems
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1794)

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Table of contents (24 protocols)

  1. Front Matter

    Pages i-xii
    Download chapter PDF

  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
      Download chapter PDF

    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
      Download chapter PDF

    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
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Keywords

About this book

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.

Editors and Affiliations

  • 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

Bibliographic Information

  • Book Title: Two-Hybrid Systems

  • Book Subtitle: Methods and Protocols

  • Editors: Luis Oñate-Sánchez

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-7871-7

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC, part of Springer Nature 2018

  • Hardcover ISBN: 978-1-4939-7870-0Published: 01 June 2018

  • Softcover ISBN: 978-1-4939-9310-9Published: 25 July 2019

  • eBook ISBN: 978-1-4939-7871-7Published: 31 May 2018

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XII, 355

  • Number of Illustrations: 8 b/w illustrations, 50 illustrations in colour

  • Topics: Protein Science

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