Yeast two-hybrid (Y2H) screening is the classical high throughput screening technology for the study of protein-protein interactions. It was first utilized by Stanley Fields and Ok-Kyu Song over 20 years ago. It is now still widely used for the initial identification of potential protein interactors. The major advantages of this technique including: 1. it is cost-effective as the technique does not require special equipments; 2. the screening is highly customizable and sensitive; 3. Y2H is an in-vivo assay, and yeast, as a eukaryotic organism, supports post translational modification and correct folding of eukaryotic proteins, which together enables discovery of physiologically relevant protein interactions.
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This technology is based on the fact that the activating domain (AD) and binding domain (BD) of a transcription factor can be separated and still function when they get into close proximity. Y2H screening is therefore designed to use a BD fused bait protein to screen against a library of AD fused prey proteins. The bait is usually a known protein that the researcher tries to identify new interacting partners with. The prey is usually a library of proteins to be screened from. When the bait protein binds to the prey protein, transcription of the specially designed reporter genes occurs and allows selection of this positive clone. Profacgen use modified yeast strains which incorporates several reporter genes to reduce false positives. We also use different GAL4 binding elements to increase sensitivity of the assay.
Profacgen offers a wide range of ready-to-use prey protein cDNA libraries. We also can custom develop homogenized cDNA libraries according to your specific requirements. Screening and validation process can be done in as short as two months! We also provide co-immunoprecipitation service to validate your result from the Y2H screening.
See more details on the principle and protocol of yeast two hybrid screening on our website. See more…
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