Spot plate assay to test the complementation of BW25113 pgpBybjGlpp::kan sensitivity to EDTA. Cells were transformed with the indicated plasmids encoding IPTG-inducible His-UppP (H-UppP), UppP, PgpB-his (PgpB-h), YgjG, or LpxT or control plasmids encoding mCherry or mKO. Plasmids encoding PgpB-his and YbjG complemented the sensitivity to EDTA with or without IPTG, though in the case of YbjG, overexpression caused slight toxicity; hence, the complementation worked better without IPTG. Plasmids encoding LpxT, UppP, or His-UppP failed to complement sensitivity to EDTA. Cells were plated in LB-Agar medium with the indicated additives and incubated for 40 h at 37 C before imaging. EDTA was added at 2 mM and IPTG at 0.1 mM. Images shown. Three colonies from each strain were tested twice; representative images are shown.
