Yeast One Hybrid Service

The yeast one-hybrid system is a technique for studying the interaction between DNA and proteins in yeast cells by observing the expression status of reporter genes in yeast cells. The yeast one-hybrid system overcomes the limitations of in vivo studies by identifying proteins stably bound to DNA, thereby screening the gene sequence encoding the protein that interacts with the target sequence directly from the DNA library, without the need to isolate and purify the protein.

Creative Proteomics offers yeast one-hybrid systems for verifying DNA-protein interactions, finding protein molecules that interact with target DNA fragments, and thus searching for novel transcription factors.

Application of Yeast One-Hybrid Technology

Currently yeast one-hybrid systems are widely used in the fields of animal genetic engineering, plant genetic engineering, medicine and pharmacology. In the study of DNA-protein interactions it is possible to:

• Identification of the presence of interactions between known DNA and known proteins
• Isolation of new proteins bound to target cis-regulatory elements or other short DNA binding sites
• Accurate localization of DNA binding sites with proven interactions and analysis of the DNA binding domains of proteins

Advantages of Yeast One-Hybrid Technology

• Directly identify proteins that bind to cis-acting elements, and also filter directly from the library to the coding sequence of the protein without the need for operational steps such as protein isolation and purification.
• Yeast is a model eukaryotic species and studies in vivo are closer to the regulation of gene expression in eukaryotes.
• The proteins interacting with DNA detected by yeast monohybridization are in their natural conformation, avoiding the shortcomings of in vitro studies.

Principle of Yeast Monohybrid Technique

Transcription initiation of eukaryotic genes requires the involvement of transcription factors. Transcription factors usually consist of a DNA-specific binding domain and one or more activation domains that regulate protein interactions, the DNA-bindingdomain (BD) and the activationdomain (AD). The DNA-binding domain of GAL4, which is used in yeast monohybrid systems, is a typical transcription factor.The DNA-binding domain of GAL4 is near the carboxy terminus and contains several zinc finger structures that activate the upstream activation site (UAS) of yeast galactosidase. In turn, the transcriptional activation structural domain can interact with RNA polymerase or transcription factor TFIID to enhance the activity of RNA polymerase. In this process, the DNA-binding structural domain and the transcription-activating structural domain can function completely independently. Accordingly, we can replace the DNA-binding structural domain of GAL4 with the library protein-encoding gene. When the expressed protein can interact with the target gene, it can activate RNA polymerase through the transcriptional activation structural domain and initiate transcription of downstream reporter genes.