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Liquid-Phase Yeast Two Hybrid assay
Our optimized protocol for liquid-phase Y2H in 96-well plates permits rapid screening of protein-protein interactions.
Our liquid assay is based on the LexA Interaction trap/two-hybrid system developped in the laboratory of Roger Brent (Golemis et al., 2001).
We have optimized the spacer sequences and the mating/culture conditions to increase the sensitivity of detection of the Beta-Galactosidase activity.
We apply it to identify:
- Key residues of a given interaction by alanine scanning & random mutagenesis,
- Potential post-translational modifications required to form/break interactions using site-directed (mimicking) mutations,
- Interacting domains of multi-domain proteins by the use of truncations,
- Interacors of a multi-protein macrocomplex by cross-mating.