Figure 4

teg58 effects l-arginine biosynthesis. (A) Northern blot analysis for various teg58 derivative strains with argG, and argH probes representing genes involved in arginine biosynthesis pathway. A total of 10 μg of cellular RNA isolated from the post-exponential phase of growth was loaded onto each lane. Ethidium bromide-stained gel used for blotting, showing 16S and 23S rRNA bands, is shown as the loading control. The relative band intensity of various lanes was scanned by Image J software (NIH) and the statistically significant was calculated considering the wild type band intensity set as 100. Original blots are presented in Supplementary Material files Fig. X3A–C. (B,C) Growth analysis in CDM containing 1% glucose without supplemented l-arginine for various isogenic strains as indicated. Growth was performed for 48 h at 37 °C in an incubator shaker (B; aerobic) and incubated in filled and sealed 50 ml screw capped tubes (C; anaerobic). (D) Determination of comparative intracellular l-arginine, l-citrulline, and l-ornithine concentrations of the wild type and the teg58 mutant of JE2 strains grown under aerobic conditions in TSB by LC–MS/MS mass spectroscopy. The asterisks indicate statistical significance between wild-type and isogenic mutant or various strains, determined using Student t-test (*, P < 0.05; **, P < 0.005; *** or ****, P < 0.0001) or “n.s” indicate “not significant”.