The Andrew Emili and Jack Greenblatt labs at the University of Toronto have created a collection of SPA- and TAP-tagged E. coli strains intended to facilitate mapping of protein-protein interactions.  Utilizing a phage recombination system, sequence-specific PCR products encoding affinity purification tags were inserted into discrete loci of the E. coli chromosome.  857 proteins, including 198 essential and conserved proteins, were successfully tagged.  Over a quarter of these proteins were also successfully purified by the source lab.

Proteins were isolated by affinity purification and their binding partners identified using LC-MS and MALDI-TOF mass spectrometry.  An extensive interaction network derived from 1000 bait ORFs was published by Butland et al. (2005).  Data generated from this study identified a reliable network of functionally diverse protein complexes. These data offer insight into the function of uncharacterized proteins and outline the topological organization of the bacterial interactome.  Knowledge of physical interactions mediated by conserved, essential bacterial proteins should facilitate the design of broad-range antimicrobials.

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