Two-Hybrid Systems

Methods and Protocols

  • Paul N. MacDonald

Part of the Methods in Molecular Biology book series (MIMB, volume 177)

Table of contents

  1. Front Matter
    Pages i-xii
  2. General Methods

    1. Front Matter
      Pages 1-1
    2. Stanley Fields, Paul L. Bartel
      Pages 3-8
    3. Lawrence W. Bergman
      Pages 9-14
    4. Michael Saghbini, Denise Hoekstra, Jim Gautsch
      Pages 15-39
    5. Philip James
      Pages 41-84
    6. Robin A. Woods, R. Daniel Gietz
      Pages 85-97
    7. Alyson Byrd, René St-Arnaud
      Pages 107-119
  3. False Positives

    1. Front Matter
      Pages 121-121
    2. Ilya G. Serebriiskii, Erica A. Golemis
      Pages 123-134
  4. Applications

  5. Alternative Strategies

    1. Front Matter
      Pages 219-219
    2. Xin-Hua Feng, Rik Derynck
      Pages 221-239
    3. Mary Kate Alexander, Brenda D. Bourns, Virginia A. Zakian
      Pages 241-259
    4. Phyllis S. Goldman, Anthony J. DeMaggio, Richard H. Goodman, Merl F. Hoekstra
      Pages 261-270
    5. Björn Sandrock, Franck Tirode, Jean-Marc Egly
      Pages 271-289
    6. Zbigniew Dominski, William F. Marzluff
      Pages 291-318
  6. Back Matter
    Pages 329-336

About this book


The yeast two-hybrid system is one of the most widely used and productive techniques available for investigating the macromolecular interactions that affect virtually all biological processes. In Two-Hybrid Systems: Methods and Protocols, Paul N. MacDonald has assembled a collection of these powerful molecular tools for examining and characterizing protein-protein, protein-DNA, and protein-RNA interactions. The techniques range from the most basic (introducing plasmids into yeasts, interaction assays, and recovering the plasmids from yeast) to the most advanced alternative strategies (involving one-hybrid, split two-hybrid, three-hybrid, membrane recruitment systems, and mammalian systems). Methods are also provided for dealing with the well-known problems of artifacts and false positives and for identifying the interacting partners in important biological systems, including the Smad and nuclear receptor pathways. To ensure ready reproducibility and robust results, each technique is described in step-by-step detail by researchers who employ it regularly.
Comprehensive and highly practical, Two-Hybrid Systems: Methods and Protocols not only reveals how the great variety of plasmid vectors and approaches may be optimally deployed, but also quickly empowers novices to establish two-hybrid systems in their laboratories, and experienced researchers to expand their repertoire of techniques.

Editors and affiliations

  • Paul N. MacDonald
    • 1
  1. 1.Department of Pharmacology, School of MedicineCase Western Reserve UniversityCleveland

Bibliographic information

  • DOI
  • Copyright Information Humana Press 2001
  • Publisher Name Humana Press
  • eBook Packages Springer Protocols
  • Print ISBN 978-0-89603-832-5
  • Online ISBN 978-1-59259-210-4
  • Series Print ISSN 1064-3745
  • Series Online ISSN 1940-6029
  • Buy this book on publisher's site
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