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A Thiamine-Regulatable Epitope-Tagged Protein Expression System in Fission Yeast

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Recombinant Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 824))

Abstract

Schizosaccharomyces pombe, the fission yeast, has been a popular and useful model system for investigating the mechanisms of biological processes for a long time. To facilitate purification, localization, and functional analysis of gene products, a wide range of expression vectors have been developed. Several of these vectors utilize the inducible/repressible promoter systems and enable the episomal expression of proteins as fusion proteins with epitope tags attached to their N terminus or C terminus.

This chapter provides a detailed protocol for expression of the epitope-tagged proteins from thiamine-regulatable nmt promoter in fission yeast. The yeast culture conditions and procedures for yeast transformation, expression induction, preparation of whole-cell extracts, and analysis of epitope-tagged protein expression by Western blotting are described.

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Acknowledgments

The author would like to thank Iain Hagan and members of the Hagan laboratory for encouragement, advice, and support and members of the Sedman’s group for discussions. This work was supported by Tartu University (baseline funding grant).

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Authors and Affiliations

  1. Institute of Molecular and Cell Biology, University of Tartu, Tartu, Estonia

    Tiina Tamm

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  1. Tiina Tamm
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Correspondence to Tiina Tamm .

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Editors and Affiliations

  1. Dept. Chemistry & Physics, Arkansas State University, Jonesboro, University Loop E. 504, State University, 72401, Arkansas, USA

    Argelia Lorence

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Tamm, T. (2012). A Thiamine-Regulatable Epitope-Tagged Protein Expression System in Fission Yeast. In: Lorence, A. (eds) Recombinant Gene Expression. Methods in Molecular Biology, vol 824. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-433-9_22

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  • DOI: https://doi.org/10.1007/978-1-61779-433-9_22

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-432-2

  • Online ISBN: 978-1-61779-433-9

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