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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Mol Microbiol. 2014 Jul 6;93(3):415–425. doi: 10.1111/mmi.12665

Fig. 1. Overproduction of CupB5 causes mucoid conversion in PAO1.

Fig. 1

(A) Organization of the cupB1-cupB6 gene cluster (www.pseudomonas.com), and location of the transposon insertion site in PAO1-VE22. (B) Alginate production, measured using the carbazole assay, and colony phenotypes (M, mucoid; NM, nonmucoid) were assayed following transformation of P. aeruginosa strain PAO1 with plasmid HERD20T or variants expressing HA-CupB4-H6, HA-CupB5-H6, or HA-CupB6-H6, and overnight growth of colonies on PIA plates (carbenecillin 300 μg ml−1, 0.1% L-arabinose) at 37°C. Values are averages ± SEM (n=3). (C) Cell lysates of PAO1 carrying appropriate plasmids were electrophoresed on SDS polyacrylamide gels, and Western-blots using an anti-HA monoclonal antibody were used to detect HA-CupB4-H6, HA-CupB5-H6, or HA-CupB6-H6. The rightmost lane shows that HA cross-reactive proteins were absent in PAO1 transformed with the pHERD20T empty-vector.