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Manipulating the Yeast Genome: Deletion, Mutation, and Tagging by PCR

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Yeast Genetics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1205))

Abstract

Saccharomyces cerevisiae is an ideal model eukaryotic system for the systematic analysis of gene function due to the ease and precision with which its genome can be manipulated. The ability of budding yeast to undergo efficient homologous recombination with short stretches of sequence homology has led to an explosion of PCR-based methods to delete and mutate yeast genes and to create fusions to epitope tags and fluorescent proteins. Here, we describe commonly used methods to generate gene deletions, to integrate mutated versions of a gene into the yeast genome, and to construct N- and C-terminal gene fusions. Using a high-efficiency yeast transformation protocol, DNA fragments with as little as 40 bp of homology can accurately target integration into a particular region of the yeast genome.

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Acknowledgements

We are indebted to participants of the Cold Spring Harbor Yeast Genetics and Genomics Course for their insights into yeast cell manipulation. We thank members of the Jaspersen lab for comments on the manuscript. S.L.J. is supported by the Stowers Institute for Medical Research and the American Cancer Society (RSG-11-030-01-CSM).

Author information

Authors and Affiliations

  1. Stowers Institute for Medical Research, 1000 E. 50th Street, Kansas City, MO, 64110, USA

    Jennifer M. Gardner & Sue L. Jaspersen

  2. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS, 66160, USA

    Sue L. Jaspersen

Authors
  1. Jennifer M. Gardner
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  2. Sue L. Jaspersen
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Corresponding author

Correspondence to Sue L. Jaspersen .

Editor information

Editors and Affiliations

  1. Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, Virginia, USA

    Jeffrey S. Smith

  2. Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, Virginia, USA

    Daniel J. Burke

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Gardner, J.M., Jaspersen, S.L. (2014). Manipulating the Yeast Genome: Deletion, Mutation, and Tagging by PCR. In: Smith, J., Burke, D. (eds) Yeast Genetics. Methods in Molecular Biology, vol 1205. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1363-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-1363-3_5

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1362-6

  • Online ISBN: 978-1-4939-1363-3

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