Membrane System Yeast Two-Hybrid (Y2H) Verification Service

Based on the traditional yeast two-hybrid system, DUALmembrane technology cleverly uses the split-ubiquitin system to screen protein interactions. The split-ubiquitin system (split-ubiquitin) can be artificially divided into N-terminal (Nub) and C-terminal (Cub). Under normal circumstances, Nub and Cub still have a high affinity, showing the function of wild-type ubiquitin, that is, still playing the signal function of degrading proteins.

How is the ubiquitin system used in yeast two-hybrid research? First, the 3rd isoleucine of ubiquitin Nub was artificially mutated to glycine (NubI was mutated to NubG). In this way, the affinity with Cub is greatly reduced, avoiding the possibility of self-association or proximity between Cub and Nub. Secondly, fusion Cub and synthetic transcription activator (LexA-VP16) into a fusion protein Cub-LexA-VP16. Therefore, under normal conditions, NubG does not bind to Cub, UAS cannot recognize isolated ubiquitin, and transcriptional activators will not be cut off. Finally, the protein to be detected is fused with NubG and Cub to form bait fusion protein (X-Cub-LexA-VP16) and prey fusion protein (Y-NubG). If bait and prey interact, it will prompt NubG and Cub to approach each other, leading to the dissociation of LexA-VP16, which will be recognized by UAS after entering the nucleus, thereby activating the transcription of the reporter gene.

Service introduction

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

Experiment process

• Bioinformatics analysis of bait protein and prey protein
• Develop experimental protocol
• Construction of bait expression vector and prey expression vector
• Detect the bait function and select the bait carrier
• Detect the working concentration of 3-AT
• Co-conversion verification
• Make a display map
• Organize data and make detailed reports

Material requirements

• Decoy sequence
• Templates and libraries

Delivery result

• Raw data
• Result picture
• Positive clones

Project cycle

It takes about 15 working days to complete yeast two-hybrid verification process of membrane 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.