BARCODED ORFEOME: SYSTEM FOR HIGH-THROUGHPUT PROTEIN-PROTEIN INTERACTION MAPPING

Here we describe two strain resources. The first resource is a collection of E. coli strains bearing uniquely barcoded BFG-Y2H compatible expression clones enabling BFG-Y2H interaction mapping for yeast proteins at proteome scale. The second resource is a collection of E. coli strains bearing uniquely barcoded BFG-Y2H compatible expression clones enabling BFG-Y2H interaction mapping for human proteins at proteome scale.Barcoded human and yeast ORFeomes provide a reusable resource for repeated screening of protein-protein interactions across many different conditions at genome scale. Conditions may include, but are not limited to, biological stimuli such as DNA damage, small molecules, as well as temperature and pH environments that modulate protein-protein interactions by affecting their physiochemical properties.Together with a next-generation sequencing instrument and a flow cytometry instrument, these clone resources enable genome scale dynamic interactomes to be mapped for yeast and human proteomes. BFG-Y2H mapping can be carried out at a fraction of a cost needed for alternative methods, while maintaining a similar quality. Where these resources of barcoded expression plasmids are used, BFG-Y2H does not require the robotics instrumentation needed for alternative similar-scale Y2H screening approaches.