Yeast Three-Hybrid (Y3H) for Protein Interaction Identification

Protein-protein interactions are closely associated with many biological processes, such as transcription, DNA modifications, and transcriptional regulations. Exploring the interactions between macromolecules contributes to revealing the molecular mechanisms of life processes. At Creative Biolabs, our experienced team of experts has built a reputation for providing cost-effective yeast three-hybrid (Y3H) services with fast turnaround times. Our comprehensive services range from initial experimental design to final analysis of results, helping you make breakthroughs in complex protein interactions.

Background of Y3H

Yeast hybrid systems are widely used due to their convenience and low cost. Based on these systems, many methods have been developed to analyze protein-protein, protein-DNA, and protein-RNA interactions. For example, the yeast two-hybrid system (Y2H) plays a crucial role in studying protein-protein interactions. Y3H, an extension of the Y2H, is a powerful tool for studying more complex macromolecular interactions involving three components.

Principles of Y3H

Y2H system is a promising tool for detecting protein interactions. In addition to the case of two proteins interacting, it is also common for a third protein to stabilize the binding between the two partners. The Y3H system has gradually emerged as an attractive strategy to explore such complex protein complexes by introducing a new bait vector, pBridge, which allows to insertion of two exogenous proteins at the same time. In this system, a bait protein is expressed with a DNA binding domain (BD) and then the intermediate protein, and the prey protein is fused with the transcriptional activation domain (AD). The Pmet25 promoter in the pBridge vector only can be expressed in the medium lacking methionine. Thus, the interaction between bait protein and prey protein through the intermediate protein is identified in the medium lacking methionine after BD and AD vectors co-transformed into the yeast strain.

Fig.1 The yeast three-hybrid system vectors. (Sandrock, 2001)

Applications of Y3H

Similar to the Y2H system, the Y3H system can be applied in various research fields.

Investigating more complex interactions between three proteins.
Investigate the interaction between two proteins and a third protein by bringing the two hybrid proteins into proximity with each other by a third protein.
The promoting or inhibiting effect of the third protein on the other two proteins was explored by the growth state of the colony.
Analysis of polyprotein-RNA complexes.
Screening for the interactions between small molecule drugs and protein interaction.

Features of Y3H

The Y3H system is easy to operate.
The third component is diverse and can be a protein, RNA, or small molecule.
Applicable to characterize protein-protein and protein-RNA interaction.
Effective detection of weak interactions between RNA/ protein and protein.

As a global company that places great emphasis on the fast delivery of solutions and cost-effective service, Creative Biolabs is committed to providing customized Y3H services for your specific project needs. Please contact us to tell us your needs, we will offer you flexible and innovative solutions and the highest quality Y3H services to help you solve the problems you encounter during your research.

Reference

Sandrock, B.; et al. Three-hybrid screens: inducible third-party systems. Methods in molecular biology (Clifton, N.J.). 2001, 177: 271-89.

All listed services and products are For Research Use Only. Do Not use in any diagnostic or therapeutic applications.

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