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Options and Considerations When Using a Yeast One-Hybrid System

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Two-Hybrid Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1794))

Abstract

Comprehensive mapping of protein–DNA interactions is essential to uncover the mechanisms involved in gene regulation. However, the data generated has been sparse given the number of regulatory elements and transcription factors (TFs) encoded in the genomes of metazoan organisms. Yeast one-hybrid (Y1H) assays provide a powerful “DNA-centered” method, complementary to “TF-centered” methods such as chromatin immunoprecipitation, to identify the TFs that can bind a DNA sequence of interest. Here, we present different technical variations that should be considered when using a Y1H system, including the type of DNA sequence to test, source of TF clones, as well as types of vectors and screening format. Finally, we discuss limitations of the assay and future challenges.

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Acknowledgments

We thank Kok Ann Gan for critically reading the manuscript. This work was supported by NIH grant GM114296 to J.I.F.B. J.S. was supported by the NIH HTP grant 5T32HL007501-34.

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Authors and Affiliations

  1. Department of Biology, Boston University, Boston, MA, USA

    Jared A. Sewell & Juan I. Fuxman Bass

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  1. Jared A. Sewell
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  2. Juan I. Fuxman Bass
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Correspondence to Juan I. Fuxman Bass .

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Editors and Affiliations

  1. Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM)–Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Campus Montegancedo UPM, Madrid, Spain

    Luis Oñate-Sánchez

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Sewell, J.A., Fuxman Bass, J.I. (2018). Options and Considerations When Using a Yeast One-Hybrid System. In: Oñate-Sánchez, L. (eds) Two-Hybrid Systems. Methods in Molecular Biology, vol 1794. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7871-7_8

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  • DOI: https://doi.org/10.1007/978-1-4939-7871-7_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7870-0

  • Online ISBN: 978-1-4939-7871-7

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