Growth phenotypes of gain-of-function mutants.

Single colonies of E. coli MJF465 containing the different pB10b derivatives were used to inoculate LB broth containing ampicillin. Cultures were grown overnight at 37°C with shaking at 200 rpm and diluted 1:200 in fresh LB broth containing ampicillin. Cultures were incubated at 37°C with shaking at 200 rpm for 2 h. The expression of mscS was subsequently induced by the addition of 1 mM IPTG, and cultures were incubated further. Throughout the time course of the growth experiments, samples were taken every hour for OD₆₀₀ measurements. All growth experiments were performed in three independent biological repeats.