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Vækst-baserede Bestemmelse og biokemisk Bekræftelse af genetiske Krav til proteinnedbrydning i<em> Saccharomyces cerevisiae</em
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JoVE Journal Biology
Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae

Vækst-baserede Bestemmelse og biokemisk Bekræftelse af genetiske Krav til proteinnedbrydning i<em> Saccharomyces cerevisiae</em

DOI:
10.3791/52428-v

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10:57 min

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February 16, 2015

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, , ,
1Department of Biology,Ball State University, 2Division of Nephrology,Cincinnati Children’s Hospital

Chapters

  • 00:05Title
  • 01:44Protein Degradation Mutant Identification Using a Yeast Growth Assay
  • 05:06Biochemical Confirmation of Protein Degradation Defects
  • 08:29Results: Ubiquitin Ligase-dependent Degradation of an Unstable Protein Fused to a Metabolic Reporter
  • 10:18Conclusion

Summary

Automatic Translation

Automatic Translation

This article describes a yeast growth-based assay for the determination of genetic requirements for protein degradation. It also demonstrates a method for rapid extraction of yeast proteins, suitable for western blotting to biochemically confirm degradation requirements. These techniques can be adapted to monitor degradation of a variety of proteins.

Tags

Keywords: Protein DegradationSaccharomyces CerevisiaeGrowth-based AssayReporter EnzymeDeg1-Sec62-His3 FusionGenetic RequirementsBiochemical ConfirmationProtein ExtractionWestern BlottingEndoplasmic Reticulum Translocon

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