Dergi
/
Genetik
/
Gene-targeted Random Mutagenesis to Select Heterochromatin-destabilizing Proteasome Mutants in Fission Yeast
JoVE Journal
Genetik
Bu içeriği görüntülemek için JoVE aboneliği gereklidir.  Oturum açın veya ücretsiz deneme sürümünü başlatın.
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

•

07:18 min

•

May 15, 2018

•

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

Bölümler

  • 00:04Başlık
  • 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

Özet

Otomatik Çeviri

Otomatik Çeviri

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.

Etiketler

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

Article

Embed

ÇALMA LİSTESİNE EKLE

Usage Statistics

İlgili Videolar

Optogenetic Random Mutagenesis Using Histone-miniSOG in C. elegans

High-Throughput Robotically Assisted Isolation of Temperature-sensitive Lethal Mutants in Chlamydomonas reinhardtii

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Using a Fluorescent PCR-capillary Gel Electrophoresis Technique to Genotype CRISPR/Cas9-mediated Knockout Mutants in a High-throughput Format

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

CRISPR/Cas9 Gene Editing to Make Conditional Mutants of Human Malaria Parasite P. falciparum

Mating-based Overexpression Library Screening in Yeast

Characterizing Histone Post-translational Modification Alterations in Yeast Neurodegenerative Proteinopathy Models

A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe

Enhanced Yeast One-hybrid Screens To Identify Transcription Factor Binding To Human DNA Sequences

Read Article