Figure 4
Phenotypic and metabolomic characterization of PtUMPS knockouts. (A) Spot plating assays of wild type (WT), \(\Delta\)UMPS1 and \(\Delta\)UMPS2 strains on L1 solid media alone, L1 supplemented with uracil, L1 supplemented with 5-FOA, or L1 supplemented with both uracil and 5-FOA. Indicated dilutions are relative to the initial concentration. (B) Liquid growth curves of wild type (WT), \(\Delta\)UMPS1 and \(\Delta\)UMPS2 strains in L1 liquid media alone, or supplemented with uracil or 5-FOA or both. Data points are the mean of three independent replicates, with error bars representing the standard error of the mean. (C) Orotate concentrations were measured by LC-MS from cultures grown with and without uracil supplementation. Bars represent mean values and error bars represent standard deviation for three biological replicates. Individual data points are represented as colored dots. Statistical confidence level was calculated by one-sided t test. p < 0.001 is indicated by an asterisk. (D) Bar graph showing percent plasmid retention in the \(\Delta\)UMPS1 and \(\Delta\)UMPS2 strains harbouring various PtUMPS constructs after 14 days of outgrowth. Bars represent the mean ratio of colonies on selective L1 + nourseothricin versus non-selective L1 plates from three independent replicates, with error bars representing the standard error of the mean.
