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. 2012 Jan;194(2):292–302. doi: 10.1128/JB.05821-11

Fig 1.

Fig 1

ZapA overproduction delocalizes FtsZ and ZapB. Combined phase-contrast and fluorescence microscopy images showing the localization of ZapB and FtsZ when ZapA is overproduced. Cells were grown at 30°C in M9 minimal medium supplemented with glucose (0.2%) and Casamino Acids (0.1%). Expression of ZapB-GFP from plasmid pEG3a, ZapB-mCherry from plasmid pEG60, FtsZ-GFP from plasmid pEG12, and ZapA from plasmids pEG58 and pEG59 was induced as described in Materials and Methods. (A) Phase-contrast image of living cells of strain MC1000/pEG58 (Plac::zapA) (+IPTG) and MC1000/pEG59 (PBAD::zapA) (+Ara) over time. NI, no inducer. (B) Phase-contrast images (a, b, and c) and FtsZ-GFP localization (a′, b′, and c′) in living cells of strain MC1000/pEG59 (PBAD::zapA)/pEG12 (Plac::ftsZ::gfp) 2 h after ZapA induction with 0.2% Ara (b and c) or without the inducer (a). ZapA levels are 8-fold higher than under wt conditions. (C) Phase-contrast (a, b, and c) images and ZapB-GFP localization (a′, b′, and c′) in living cells of strain MC1000/pEG58 (Plac::zapA)/pEG3a (PBAD::zapB::gfp) 2 h after ZapA induction with 500 μM IPTG (b) or 1 mM IPTG (c) or without the inducer (a). ZapA levels are 7-fold (500 μM IPTG) and 12-fold (1 mM IPTG) higher than under wt conditions. (D) Phase-contrast images (a, b and c), FtsZ-GFP localization (a′, b′, and c′), and ZapB-mCherry localization (a″, b″, and c″) in living cells of strain DH5α/pEG59 (PBAD::zapA)/pEG12 (Plac::ftsZ::gfp)/pEG60 (Ptol::zapB::mCherry) 2 h after ZapA induction with 0.2% Ara (b and c) or without the inducer (a). ZapA levels are 8-fold higher than under wt conditions. (E) Localization of YFP-ZapA in living cells of strain MC1000/pNG53 (Plac::yfp::zapA) 2 h after induction with 200 μM IPTG (b), 500 μM IPTG (c), or 1 mM IPTG (d) or without the inducer (a). Scale bar, 2 μm.