A novel genetic system to detect protein–protein interactions

@article{Fields1989ANG,
  title={A novel genetic system to detect protein–protein interactions},
  author={Stanley Fields and Ok‐kyu Song},
  journal={Nature},
  year={1989},
  volume={340},
  pages={245-246}
}
PROTEIN-protein interactions between two proteins have generally been studied using biochemical techniques such as crosslinking, co-immunoprecipitation and co-fractionation by chromatography. We have generated a novel genetic system to study these interactions by taking advantage of the properties of the GAL4 protein of the yeast Saccharomyces cerevisiae. This protein is a transcriptional activator required for the expression of genes encoding enzymes of galactose utilization1. It consists of… 

Yeast two-hybrid assay for studying protein-protein interactions.

  • A. Osman
  • Biology
    Methods in molecular biology
  • 2004
This chapter presents an outline for the GAL4-based yeast two-hybrid system with a detailed description of the vectors, host cells, and methods for detection and verifying protein interactions.

Using the yeast two-hybrid system to identify protein-protein interactions.

Protocols to test interactions between two individual proteins and to look for novel interacting partners by screening a single protein or domain against a library of other proteins using a GAL4 based yeast two-hybrid system are described.

A new LexA-based genetic system for monitoring and analyzing protein heterodimerization in Escherichia coli

A new method for specifically detecting the heterodimerization of two heterologous proteins in the bacterium Escherichia coli is developed, based on the simultaneous use of protein fusions with an altered specificity and a wild-type LexA repressor DNA-binding domain.

Generation of a yeast two-hybrid strain suitable for competitive protein binding analysis.

The yeast two-hybrid (Y2H) system has been broadened and applied for further characterization of protein-protein interactions, and the URA3 gene is replaced in the standard strain L40c, aimed to create a strain still exploitable for Y2H screens, but additionally allowing the simultaneous expression of a third protein.

Mapping Protein-Protein Interaction Using High-Throughput Yeast 2-Hybrid.

This chapter provides a step-by-step protocol for the Y2H experimentation preceded by a materials listing while simultaneously including key notes throughout the entire experimental process of biological-mechanistic and historical understandings of the steps.

Identification of interacting proteins using the yeast two-hybrid screen.

A two-hybrid yeast system has been used to successfully characterize protein interactions among signaling molecules and can be used to identify new binding partners for a protein of interest or define the interaction domains and relative affinity between two proteins known to interact.
...

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