Yeast One-Hybrid Screening
Yeast One-Hybrid technology was first developed from yeast two hybrid technology by Li et al. In 1993. Through the phenotypic detection of reporter genes, the interaction between DNA and protein was analyzed to study the regulation of gene expression in eukaryotic cells. Due to the sensitivity and reliability of yeast one hybrid method in detecting the specific interaction between specific transcription factors and cis acting elements, yeast one hybrid method has been widely used to clone specific transcription factors which are weak in content and difficult to purify by biochemical means.
Yeast One-Hybrid is an effective method for cloning transcription factor genes (cDNAs) that specifically bind to target elements, based on the principle that DNA binding proteins (transcription factors) bind to DNA cis acting elements to regulate reporter gene expression. The theoretical basis is: many eukaryotic transcriptional activators are composed of physically and functionally independent DNA binding domain (BD) and activation domain (AD), so they can construct fusion expression vectors of various genes and AD. Then we can screen out the proteins that have specific binding regions with the target elements. Theoretically, in one hybrid detection, any target element can be used to screen a protein with a specific binding region.
Application of yeast single hybridization:
- Determine if there is interaction between known DNA with proteins;
- Screening of new genes that are linked to the target cis regulatory element or other short DNA sequences;
- Identify the domain of protein binding DNA and accurately locate the protein sequence that binds to DNA.
Experiment workflow:
- Establishment and screening of Library
- Protein-protein interaction verification
- Self-activation and toxicity verification
- Yeast plasmid extraction and sequencing
Based on yeast one hybrid technology, Profacgen has established a transcription factor centered technology to identify its recognition elements. This technology can accurately, quickly, simply, and efficiently identify the elements recognized by transcription factors, and has a broad application prospect in the study of the interaction between protein and DNA elements. If you are interested in our Yeast One-Hybrid Screening service, Profacgen looks forward to cooperating with you.