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. 2018 Dec 15;47(4):1740–1758. doi: 10.1093/nar/gky1256

Figure 5.

Figure 5.

Determination of SprG2 and SprG3 internal coding sequences, with evidence that both trigger Staphylococcus aureus stasis but not death. (A) Schematic showing ‘pALCΩSprG2-STOP’ and ‘pALCΩSprG3-STOP’ strain constructs overexpressing either SprG2 or SprG3, with stop codons instead of the predicted initiation codons in order to abolish peptide production. (B) Growth kinetics in liquid cultures and (C) serial dilutions of exponentially grown HG003 bacteria containing pALC (yellow), pALCΩSprG2 or pALCΩSprG3 (blue), pALCΩSprG2-STOP (green), pALCΩSprG3-STOP1 (black), pALCΩSprG3-STOP2 (green), pALCΩSprG3-STOP3 (gray) or pALCΩSprG3-STOP4 (orange) RNA after induction with anhydrotetracycline (aTc). Error bars indicate the variations between three biological replicates. (D) Pictures of S. aureus cells labeled with a SYTO 9 fluorescent probe after LIVE/DEAD experiments demonstrate that overexpression of SprG2- and SprG3-encoded peptides induce stasis. HG003 bacteria containing pALCΩSprG1 are shown as a positive control for S. aureus death, as well as HG003 containing empty pALC treated with 70% isopropanol.