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. 2012 Nov 26;7(11):e49749. doi: 10.1371/journal.pone.0049749

Figure 4. Complementation of ΔbamB phenotype in E. coli.

Figure 4

(A) Cells expressing control plasmid (GFP) do not grow at vancomycin concentrations higher than 60 µg/ml (no cell pellets in the round bottom wells) whereas plasmids expressing wild type E. coli or Pseudomonas BamB (ecBamb and paBamB respectively) complement the ΔbamB phenotype and grow at higher vancomycin concentrations (at least 150 µg/ml). All the ecBamB mutants tested were able to complement the ΔbamB phenotype except D341A. (B) Western blots of total and detergent solubilized E. coli ΔbamB cells expressing wild type (ecBamB) or mutant (D341A) BamB from a plasmid. Equivalent cell loads (controlled by probing with BamA antibody) show that the ecBamB D341A mutant expresses at a lower level and fails to properly assemble in the membrane (see text for details).