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Reporter-based Growth Assay for Systematic Analysis of Protein Degradation
JoVE Journal
Biologia
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JoVE Journal Biologia
Reporter-based Growth Assay for Systematic Analysis of Protein Degradation
DOI:

07:47 min

November 06, 2014

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Capitoli

  • 00:05Titolo
  • 01:30Growth Kinetics in Liquid Culture Under Selective Conditions (GiLS)
  • 03:36Calculation of the Minimal Doubling Time (MDT) Using MDTcalc
  • 05:04Results: Representative Protein Degradation Analysis
  • 06:45Conclusion

Summary

Traduzione automatica

Here we describe a robust biological assay for quantifying the relative rate of proteolysis by the ubiquitin-proteasome system. The assay readout is yeast growth rate in liquid culture, which is dependent on the cellular levels of a reporter protein comprising a degradation signal fused to an essential metabolic marker.

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