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A Stringent Yeast Two-Hybrid Matrix Screening Approach for Protein–Protein Interaction Discovery

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Two Hybrid Technologies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 812))

Abstract

The yeast two-hybrid (Y2H) system is currently one of the most important techniques for protein–protein interaction (PPI) discovery. Here, we describe a stringent three-step Y2H matrix interaction approach that is suitable for systematic PPI screening on a proteome scale. We start with the identification and elimination of autoactivating strains that would lead to false-positive signals and prevent the identification of interactions. Nonautoactivating strains are used for the primary PPI screen that is carried out in quadruplicate with arrayed preys. Interacting pairs of baits and preys are identified in a pairwise retest step. Only PPI pairs that pass the retest step are regarded as potentially biologically relevant interactions and are considered for further analysis.

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Acknowledgments

J.M. Worseck, A. Grossmann, and M. Weimann contributed equally to the writing of this protocol. We would like to thank Erich Wanker (MDC-Berlin) and the members of his group for continuing support and for their contributions in developing the Y2H setup.

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Authors and Affiliations

  1. Max Planck Institute for Molecular Genetics (MPI-MG), Berlin, Germany

    Josephine M. Worseck, Arndt Grossmann, Mareike Weimann, Anna Hegele & Ulrich Stelzl

Authors
  1. Josephine M. Worseck
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  2. Arndt Grossmann
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  3. Mareike Weimann
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  4. Anna Hegele
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  5. Ulrich Stelzl
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Corresponding author

Correspondence to Ulrich Stelzl .

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Editors and Affiliations

  1. Molekulare Medizin (MDC), Max-Delbrück-Centrum für, Robert-Rössle-Str. 10, Berlin-Buch, 13125, Germany

    Bernhard Suter

  2. Max-Delbrück-Centrum-Für Molekulare Medi, Robert-Rössle-Strabe 10, Berlin, 13125, Berlin, Germany

    Erich E. Wanker

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Worseck, J.M., Grossmann, A., Weimann, M., Hegele, A., Stelzl, U. (2012). A Stringent Yeast Two-Hybrid Matrix Screening Approach for Protein–Protein Interaction Discovery. In: Suter, B., Wanker, E. (eds) Two Hybrid Technologies. Methods in Molecular Biology, vol 812. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-455-1_4

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  • DOI: https://doi.org/10.1007/978-1-61779-455-1_4

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-454-4

  • Online ISBN: 978-1-61779-455-1

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