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Study of Amyloids Using Yeast

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Amyloid Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 849))

Abstract

We detail some of the genetic, biochemical, and physical methods useful in studying amyloids in yeast, particularly the yeast prions. These methods include cytoduction (cytoplasmic mixing), infection of cells with prion amyloids, use of green fluorescent protein fusions with amyloid-forming proteins for cytology, protein purification and amyloid formation, and electron microscopy of filaments.

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Abbreviations

USA:

Ureidosuccinic acid

SD:

Synthetic dextrose (medium)

SC-Ade:

Synthetic complete minus adenine (medium)

YPAD:

Yeast extract peptone adenine dextrose (medium)

YPG:

Yeast extract peptone glycerol (medium)

GFP:

Green fluorescent protein

NiNTA:

Nickel column

TCA:

Trichloroacetic acid

TEM:

Transmission electron microscopy

TMV:

Tobacco mosaic virus

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Acknowledgments

This work was supported by the Intramural Program of the National Institute of Diabetes Digestive and Kidney Diseases. The electron diffraction method was developed by Kent Thurber and Rob Tycko (NIH, Bethesda, MD).

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Authors and Affiliations

  1. Laboratory of Biochemistry and Genetics, National Institute of Diabetes Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA

    Reed B. Wickner, Dmitry Kryndushkin, Ryan McGlinchey & Herman K. Edskes

  2. Department of Pharmacology, Uniformed Services University of the Health Sciences, Bethesda, MD, USA

    Dmitry Kryndushkin & Frank Shewmaker

Authors
  1. Reed B. Wickner
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  2. Dmitry Kryndushkin
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  3. Frank Shewmaker
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  4. Ryan McGlinchey
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  5. Herman K. Edskes
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Corresponding author

Correspondence to Reed B. Wickner .

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Editors and Affiliations

  1. School of Medicine, Dept. Psychiatry, New York University, First Ave. 550, New York, 10016, New York, USA

    Einar M. Sigurdsson

  2. Centro Nacional de Microbiología, Instituto de Salud Carlos III, Carretera Majadahonda-Pozuelo Km 2, Madrid, 28220, Spain

    Miguel Calero

  3. Inst. Química Física, Rocasolano, CSIC Madrid, C/ Serrano 119, Madrid, 28006, Spain

    María Gasset

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Wickner, R.B., Kryndushkin, D., Shewmaker, F., McGlinchey, R., Edskes, H.K. (2012). Study of Amyloids Using Yeast. In: Sigurdsson, E., Calero, M., Gasset, M. (eds) Amyloid Proteins. Methods in Molecular Biology, vol 849. Humana Press. https://doi.org/10.1007/978-1-61779-551-0_22

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  • DOI: https://doi.org/10.1007/978-1-61779-551-0_22

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-550-3

  • Online ISBN: 978-1-61779-551-0

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