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Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
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JoVE Journal Genetik
Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast

DOI:
10.3791/57499-v

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07:18 min

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May 15, 2018

•

,
1Department of Biological Sciences,Korea Advanced Institute of Science and Technology

Kapitel

  • 00:04Titel
  • 00:37Random Mutagenesis of rpt4+ by Error-prone PCR
  • 01:33Transformation of Fission Yeast by Electroporation
  • 04:01Selection of Heterochromatin-destabilizing Mutants and Checking for False Positives
  • 05:44Results: Random Mutagenesis of Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
  • 06:36Conclusion

Summary

Automatische Übersetzung

Automatische Übersetzung

This article describes a detailed methodology for random mutagenesis of a target gene in fission yeast. As an example, we target rpt4+, which encodes a subunit of the 19S proteasome, and screen for mutations that destabilize heterochromatin.

Tags

Gene-targeted Random MutagenesisHeterochromatin-destabilizingProteasome MutantsFission YeastError-prone PCRTransformation-fusion ConstructElectroporationSorbitol WashHeterochromatin Formation And Maintenance

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