Journal
/
生物学
/
Split-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions
JoVE 杂志
生物学
需要订阅 JoVE 才能查看此.  登录或开始免费试用。
JoVE 杂志 生物学
Split-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions

Split-Ubiquitin Based Membrane Yeast Two-Hybrid (MYTH) System: A Powerful Tool For Identifying Protein-Protein Interactions

DOI:
10.3791/1698-v

•

14:04 min

•

February 01, 2010

•

, , ,
1Department of Biochemistry,University of Toronto, 2Department of Molecular Genetics,University of Toronto, 3Terrence Donnelly Centre for Cellular and Biomolecular Research (CCBR),University of Toronto

Chapters

  • 00:00Title
  • 01:22Introduction
  • 01:41Selection of Bait and Appropriate MYTH System
  • 03:04Generation of tMYTH Baits by Gap Repair
  • 03:07a. Vector Preparation and Amplification of Gene of Interest
  • 05:30b. Yeast Transformation and Recovery of Bait Plasmid DNA
  • 07:03Bait Validation
  • 08:10MYTH Screening
  • 08:12a. Large Scale Yeast Transformation
  • 11:31b. Bait Dependency Test
  • 12:54Representative Results/ Outcome
  • 13:12Conclusion

Summary

自动翻译

自动翻译

MYTH allows the sensitive detection of transient and stable interactions between proteins that are expressed in the model organism Saccharomyces cerevisiae. It has been successfully applied to study exogenous and yeast integral membrane proteins in order to identify their interacting partners in a high throughput manner.

Tags

Split-ubiquitinMembrane Yeast Two-hybridProtein-protein InteractionsMYTH SystemFull-length Transmembrane ProteinsSaccharomyces CerevisiaeBait ProteinPrey ProteinsUbiquitin MoietiesArtificial Transcription FactorPseudo-ubiquitin MoleculeDeubiquitinating EnzymesReporter Gene Expression

Article

Embed

添加到播放列表

Usage Statistics

Related Videos

Imaging Protein-protein Interactions in vivo

Associated Chromosome Trap for Identifying Long-range DNA Interactions

Modified Yeast-Two-Hybrid System to Identify Proteins Interacting with the Growth Factor Progranulin

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

Identifying Protein-protein Interaction in Drosophila Adult Heads by Tandem Affinity Purification (TAP)

Reverse Yeast Two-hybrid System to Identify Mammalian Nuclear Receptor Residues that Interact with Ligands and/or Antagonists

Capture Compound Mass Spectrometry – A Powerful Tool to Identify Novel c-di-GMP Effector Proteins

Investigating Protein-protein Interactions in Live Cells Using Bioluminescence Resonance Energy Transfer

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Identifying Protein-protein Interaction Sites Using Peptide Arrays

Read Article