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Identification of proteins that interact with a protein of interest: Applications of the yeast two-hybrid system

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Abstract

The yeast two-hybrid system is a molecular genetic test for protein interaction. Here we describe a step by step procedure to screen for proteins that interact with a protein of interest using the two-hybrid system. This process includes, construction and testing of the bait plasmid, screening a plasmid library for interacting fusion proteins, elimination of false positives and deletion analysis of true positives. This procedure is designed to allow investigators to identify proteins and their encoding cDNAs that have a biologically significant interaction with your protein of interest.

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Authors and Affiliations

  1. Department of Human Genetics, University of Manitoba, Winnipeg, Manitoba, R3E 0W3, Canada

    R. Daniel Gietz, Anne Robbins & Kevin C. Graham

  2. Department of Biochemistry and Molecular Biology, University of Manitoba, Winnipeg, Manitoba, R3E 0W3, Canada

    Barbara Triggs-Raine

  3. Department of Biology, University of Winnipeg, Winnipeg, Manitoba, R3B 2E9, Canada

    Robin A. Woods

Authors
  1. R. Daniel Gietz
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  2. Barbara Triggs-Raine
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  3. Anne Robbins
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  4. Kevin C. Graham
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  5. Robin A. Woods
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Gietz, R.D., Triggs-Raine, B., Robbins, A. et al. Identification of proteins that interact with a protein of interest: Applications of the yeast two-hybrid system. Mol Cell Biochem 172, 67–79 (1997). https://doi.org/10.1023/A:1006859319926

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  • DOI: https://doi.org/10.1023/A:1006859319926

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