A key part of gene functional analysis and potential drug target discovery is an understanding of how proteins interact within the cell. The two-hybrid system is a very powerful molecular genetic tool which investigates protein-protein interactions in vivo. This system can be used to study the interaction between two proteins which are expected to interact or find proteins (prey) that interact with a protein you already have (bait).
In Creative Biolabs, we can provide many types of two hybrid services, such as yeast two-hybrid (Y2H), mammalian two-hybrid (M2H), bacterial two-hybrid (B2H) or reverse yeast two-hybrid (rYTH), for global clients to identify the role of protein-protein interactions in biological systems. By using the method offered by us, scientists are able to explain structure-function relationships among proteins and mediate the biological function of interactors for therapeutic purposes, as well as to establish large-scale protein interaction maps. Additionally, as these techniques can be applied in drug discovery and development programs, they also could be new opportunities for biomedical research. We offer services on a strictly fee-for-service basis and several major two-hybrid services are introduced as follows:
It is acknowledged that Y2H is a greatly powerful technique for studying protein-protein interactions. In the Y2H assay, the 'bait' and 'hunter' plasmids are introduced into yeast cells by transfection. In this process, the plasma membrane is disrupted to yield holes, through which the plasmids can enter. Once transfection has occurred, cells containing both plasmids are selected for by growing cells on minimal media. Only cells containing both plasmids have both genes encoding for missing nutrients, and consequently, are the only cells that will survive.
Fig.1 Yeast two-hybrid system.
The principle of M2H is analogous to the better-known Y2H. The M2H technology relies on the mammalian background in which they operate and complement the established Y2H approach. This assay opens a new avenue to study the dynamics of mammalian protein interaction networks, including the temporal, spatial and functional modulation of proteins associations.
Fig.2 Mammalian two-hybrid layout.
The B2H assay is based on the reconstitution of adenylate cyclase in Escherichia coli that was described 14 years ago. For microbiologists, it is indeed a practical and useful alternative to the use of the widely spread Y2H system for testing protein-protein interactions.
Fig.3 Bacterial two-hybrid protocol.
rY2H is an upside-down version of Y2H, in which protein interaction is lethal or toxic for the yeast cells. Yeast strain expressing URA3 gene can’t grow, so URA3 is usually chosen for negative selection.
Creative Biolabs has professional teams and labs for two-hybrid services and the efficiency and quality of our identifying and screening platforms have always been satisfying clients with highly reproducible and reliable results. Except that, our two-hybrid techniques have other predominant advantages as well.