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Center for Molecular Medicine and Genetics
Wayne State University School of Medicine
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Detroit, MI 48201


This is a preprint version of an article published in Methods in Enzymology*, Applications of Chimeric Genes and Hybrid Proteins, Part C, Protein-Protein Interactions and Genomics, eds John Abelson, Scott Emr, and Jeremy Thorner, vol 328, pp26-46, 2000

*the only definitive repository of the content that has been certified and accepted after peer review. Copyright and all rights therein are retained by Academic Press. This material may not be copied or reposted without explicit permission. Copyright 2000 Academic Press.


Interaction mating methods in two-hybrid systems

Mikhail G. Kolonin, Jinhui Zhong, and Russell L. Finley Jr.

Center for Molecular Medicine and Genetics

Wayne State University School of Medicine

Detroit, Michigan 48201

Paper in PDF format



Isolating new interactors from libraries

  • Constructing and characterizing the bait strain

    Preparing the pretransformed library and control strains

    Screening for interactors by mating

    Characterizing interactors

Testing interactions between small sets of proteins by cross-mating assay

Mapping networks of interacting proteins

  • Elaborating protein networks by reiterative interactor hunts

    Constructing new bait strains by recombination cloning

    Screening two-hybrid arrays

Exploring the functions of individual protein interactions

  • Isolating interaction mutants and suppressors

    Breaking interactions with proteins and peptides

    Isolating disruptive peptide aptamers

    Interpreting results

Concluding remarks

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