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We can even screen your transmembrane baits!
To identify the interacting partners of an integral transmembrane (TM) protein, you can take advantage of our exclusive MBmate Y2H. This is a modified Yeast Two-Hybrid technique in which the bait and its partners are expressed at the yeast's plasma membrane.
Based on a split-ubiquitin system, MBmate Y2H is the only service allowing to screen full TM baits rather than cytoplasmic, extracellular or lumenal fragments.
Your membrane protein of interest is fused to the C-terminal half of ubiquitin (Cub) linked to a transcription factor (TF), LexA-VP16. Each prey in the library is fused to the N-terminal half of ubiquitin (Nub) bearing a point mutation resulting in a low affinity for Cub. Upon interaction between the bait and a prey, Cub and Nub re-associate to form an ubiquitin molecule, which is then recognized by endogenous proteases releasing the TF. The transcription of the His3 reporter gene is activated and allows yeast cells to grow on a medium lacking histidine. The DNA of the positive clones is then sequenced and analyzed to identify the protein partners.
For a detailed explanation of the MBmate Y2H principle and the split-ubiquitin system, please ask us.
The results of your MBmate Y2H screen come fully analyzed to help you focus on the most relevant interactions. The deliverables include membrane localization test results, the list of identified protein partners, the experimental interaction domain on each protein and a confidence score. Please learn more on our deliverables here.
Hesitating between ULTImate Y2H or MBmate Y2H technology? Get full advice from our experts to choose the best method for your protein!