The Matchmaker Gold Yeast Two-Hybrid System is the most advanced and highest performing version of our highly popular Matchmaker product line for investigating protein-protein interactions. This is due to a combination of a new yeast strain (Y2H Gold), stringent reporters, easy-to-use libraries, high-level expression vectors, and SMART-based library construction kits.
Matchmaker Yeast Two-Hybrid System
In a Matchmaker GAL4-based two-hybrid assay, a bait protein is expressed as a fusion to the GAL4 DNA-binding domain (DNA-BD), while libraries of prey proteins are expressed as fusions to the GAL4 activation domain (AD; 1–2). When bait and library (prey) fusion proteins interact, the DNA-BD and AD are brought into proximity to activate transcription of four independent reporter genes (AUR1-C, ADE2, HIS3, and MEL1).
This technology can be used to:
- Identify novel protein interactions
- Confirm suspected interactions
- Define interacting domains
Why do I need 4 reporter genes?
Yeast two-hybrid systems that utilize only the HIS3 nutritional reporter to screen for protein-protein interactions often generate a high number of background colonies and false positives. Background colonies are the result of leaky expression of the nutritional reporter, whereas false positives are caused by prey proteins that, independently of the bait, have the ability to recognize and bind to sequences upstream of the reporter.
The Matchmaker Gold Yeast Two-Hybrid System is the only yeast two-hybrid system with four genuine reporters and three different bait recognition sequences. Background growth is not possible due to the potency and stability of Aureobasidin A, a novel yeast two-hybrid antibiotic that kills all non-resistant cells. False positives are reduced to a minimum because any independently binding prey protein would need to bind to all 3 recognition sequences and activate 4 different reporters.
The 4 Reporters
The system utilizes antibiotic, nutritional and colorimetric reporters.
Yeast Two-Hybrid Media Sets
Our two-hybrid yeast media sets each contain a complete set of the all the Yeast Media Pouches you need for Clontech’s Matchmaker Gold Two-Hybrid protocols.