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Related Experiment Videos

Reporter assay systems for [URE3] detection and analysis.

Andreas Brachmann1, James A Toombs, Eric D Ross

  • 1Department Biologie I, Ludwig-Maximilians-Universität, Maria-Ward-Str. 1a, 80638 München, Germany. a.brachmann@lrz.uni-muenchen.de

Methods (San Diego, Calif.)
|June 10, 2006
PubMed
Summary

New methods improve the study of the Saccharomyces cerevisiae [URE3] prion, the amyloid form of Ure2p protein. These assays offer higher stringency and flexibility for prion research.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Yeast Genetics

Background:

  • The Saccharomyces cerevisiae prion [URE3] is an amyloid form of the Ure2p protein.
  • It serves as a model for studying in vivo amyloid formation and stability.
  • The presence of [URE3] enables ureidosuccinate uptake, a common but limited assay method.

Purpose of the Study:

  • To describe novel, alternative selection methods for detecting the [URE3] prion.
  • To overcome the limitations of the traditional ureidosuccinate uptake assay.
  • To enable higher stringency and flexibility in prion research.

Main Methods:

  • Utilized the Ure2p-regulated DAL5 promoter.
  • Employed reporter genes including ADE2, URA3, kanMX, and CAN1.

Related Experiment Videos

  • Developed methods for high-stringency selection and nonselective assay of prion variants.

    • Main Results:

    • Introduced alternative assays for [URE3] presence or absence.
    • Achieved higher stringency in both positive and negative selection for the prion.
    • Enabled nonselective assay of prion variants and direct transformation of prion filaments.

    Conclusions:

    • The new methods offer significant advantages over the traditional assay.
    • These assays enhance experimental possibilities in [URE3] prion research.
    • Discussed the specific benefits and drawbacks of each newly developed assay.