Nuclear System Yeast Two-Hybrid (Y2H) Library Screening Service

Yeast two-hybrid was proposed by Fields in 1989, and its production was based on the study of the properties of eukaryotic transcription factors (especially GAL4). GAL4 includes two domains that are separated from each other but are essential for function. The DNA binding domain (BD) located in the segment of amino acid residues 1-147 of the N-terminal and the transcription activation domain (AD) located in the segment of amino acid residues of 768-881 in the C-terminal. DNA-BD can recognize and bind to the upstream activation sequence (UAS) of GAL4-responsivegene. AD is through the combination with other components in the transcriptionmachinery to initiate transcription of genes downstream of UAS. DNA-BD and AD acting separately cannot activate the transcription reaction, but when the two are sufficiently close in space, they exhibit complete GAL4 transcription factor activity and can activate the promoter downstream of UAS, so that genes downstream of the promoter can be transcribed.

Fields has established a two-hybrid system in which BD is fused with X protein, and AD is fused with Y protein. If a protein-protein complex is formed between X and Y, and the two domains of GAL4 are reconstituted, then the specific gene sequence will be activated. Transcription. Generally, the fusion protein of BD-X is called bait, X is often a known protein, AD-Y is called prey, and genes that can show the interaction between bait and prey are called reporter genes. Conversely, through the detection of the reporter gene, it can be judged whether there is an interaction between the bait and the prey.

Service introduction

Creative BioMart combines years of library construction experience to provide yeast two-hybrid (Y2H) library screening service of nuclear system.

Experiment process

• Bioinformatics analysis of bait protein and prey protein
• Formulation of experimental protocol
• Construction of bait protein expression vector
• Verification of expression vector transformed yeast, self-activation activity test, toxicity test
• Library transformation or mating
• Transfer of positive clones
• Sequencing analysis of library screening results
• Rotation verification of screening results (optional)
• Organize data and make detailed reports

Delivery result

• Detailed screening report, including sequencing results of screening results.
• Library screening picture (JPG)
• Plasmids for screening results
• (Optional) library glycerol bacteria

Project cycle

It takes about 45 working days to complete yeast two-hybrid library screening process of nuclear system.

Service process

Please let us know the details of your yeast related projects through online consultation. We will reply to you in time and provide you with a comprehensive evaluation report on your project, including the feasibility evaluation of the project and the detailed experimental plan designed by our professionals. As our condition optimization is completed, the result report and other final products will be delivered to you immediately.

Creative BioMart has accumulated many years of experience in the field of yeast research, helping our customers accelerate research in all fields using yeast as a research tool, and improve the overall success rate of the project. Please tell us your project requirements, and we will provide you with a full service from strategy design to final report. If you have any questions, please feel free to contact us.