Figure 5. Hypomorphic mutants in the PG precursor synthesis pathway rescue ΔdacA1 growth but not morphology.

(A) Indicated strains are chromosomal replacements of murA and murD murA^P122S, murA^L35F and murD^D447E respectively. Overnight cultures were plated on LB medium by spot- titering and incubated at 30°C overnight. (B) Overnight cultures of the indicated strains were subcultured 1:100 and grown at 37°C in LB medium for 1 hour, then backdiluted 1:100 in LB and incubated at 37°C for an additional 2 hours before imaging (C) The different MurA mutants were purified, and their activity was measured in vitro using a MurA assay kit which measures the inorganic phosphate released from the MurA enzymatic reaction. Statistical significance was determined by an ordinary one-way ANOVA, error bars represent the standard deviation of three replicates (raw data points also shown).