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Protein Targeting Protocols pp 101–131Cite as

Molecular Genetic Approaches I

Two-Hybrid Systems

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 88))

Abstract

The two-hybrid system is a powerful yeast-based genetic system for isolating cDNAs encoding proteins that interact with a protein of interest. First proposed by Fields and colleagues (1), the two-hybrid system has enabled investigators to isolate and characterize numerous protein-protein interactions, and to isolate novel interacting partners for many biologically important enzyme complexes, signaling proteins, and transcription factors. In some cases, the two-hybrid system has demonstrated interactions between proteins that were either too transient, or too unstable, to be detected by traditional biochemical analysis. However, the two-hybrid system can also generate numerous false-positive interactions that often complicate the analysis of results obtained with this method. This chapter will first describe the theoretical basis for the two-hybrid system, and provide a primer for understanding the yeast genetics needed for a full understanding of the method. It will then provide several detailed protocols for performing a two-hybrid screen. It will describe a battery of genetic tests required for confirming positives isolated by the method.

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Author information

Authors and Affiliations

  1. Veterans Affairs Medical Center, USA

    Graeme B. Bolger

  2. Departments of Medicine (Hematology/Oncology) and Oncologic Science, Huntsman Cancer Institute, University of Utah, Salt Lake City, UT

    Graeme B. Bolger

Authors
  1. Graeme B. Bolger
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Editor information

Editors and Affiliations

  1. Hannah Research Institute, Scotland, UK

    Roger A. Clegg

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© 1998 Humana Press Inc.

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Bolger, G.B. (1998). Molecular Genetic Approaches I. In: Clegg, R.A. (eds) Protein Targeting Protocols. Methods in Molecular Biology™, vol 88. Humana Press. https://doi.org/10.1385/0-89603-487-9:101

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  • DOI: https://doi.org/10.1385/0-89603-487-9:101

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-487-7

  • Online ISBN: 978-1-59259-572-3

  • eBook Packages: Springer Protocols

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