Yeast Three-hybrid (Y3H)

Yeast Three-hybrid (Y3H)

Introduction

In cells, RNA-protein interactions play a vital role in various biological processes including virus reproduction. For example, in eukaryotes, cells need large amounts of RNA-binding proteins to control the mRNA content. In order to study these life activities in depth, scientists have developed many biochemical or genetic methods.

The yeast three-hybrid system (Y3H) is a derivative of the yeast two-hybrid (Y2H), a powerful tool capable of analyzing RNA-protein interactions, and usually consists of three chimeric components. The first hybrid protein consists of an RNA binding protein (RBD) fused to a DNA binding domain (DBD). The second hybrid protein molecule contains a second RNA binding protein fused to the transcriptional activation domain (AD). The third hybrid part is an RNA molecule, which can provide two specific RNA targets for the RNA binding proteins to bridge the two fusion proteins. When this tripartite component is formed at a promoter, the reporter gene will be turned on, and the expressed reporter products can be identified by simple biochemical or phenotypic analysis. One of the major advantages of this method is the ability to analyze RNA-protein interactions in an in vivo environment.

The yeast three-hybrid system designed by SenGupta et al. and Putz et al.Figure 1. The yeast three-hybrid system designed by SenGupta et al. and Putz et al. (Martin, F. 2012)

Services

Creative Proteomics is a professional biotechnology company that recruited numerous genetics and biochemistry experts, and established various high-quality technology platforms, including the advanced yeast three-hybrid system. We have accumulated more than 20 years of experience on Y2H and Y3H, helping thousands of researchers from all over the world complete related projects.

In addition, our experts can customize experiments according to your needs, for example, increase the expression level of AD-protein to compensate for weak binders and increase the sensitivity of this approach; introduce selection markers such as ADE2 into the RNA encoding plasmid; use 5-FOA for counter-selection; use new strains such as YBZ1, etc.

Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.

Applications

  • Search for protein chaperones of a known RNA sequence in cDNA libraries
  • Search the RNA libraries for natural RNA partners or ligands of known RNA binding proteins
  • Discovery of multiprotein-RNA complex
  • Investigate potential RNA-protein interactors
  • Mutation analysis of interacting RNA and protein

Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. The yeast three-hybrid platform we constructed has the characteristics of high quality and efficiency, and the data obtained can be directly used for paper publication. Our one-stop service aims to save customers time and money.    

References

  1. Martin, F. Fifteen years of the yeast three-hybrid system: RNA-protein interactions under investigation. Methods. 2012, 58(4).
  2. Cottier, S.; et al. The yeast three-hybrid system as an experimental platform to identify proteins interacting with small signaling molecules in plant cells: potential and limitations. Front. Plant Sci. 2011.
* This service is for RESEARCH USE ONLY, not intended for any clinical use.

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