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Effect of the addition of c-di-GMP on CtrA phosphorylation in a phosphorelay assay. The presence of c-di-GMP causes a change from phosphorylation of CtrA to dephosphorylation of CtrA∼P. Samples were removed at the indicated time intervals (given in minutes); after 15 min, the reaction mixture was split, and c-di-GMP was added to one of two tubes.
