TABLE 2.
Phenotypes of ECL3501 expressing mutant cpx operons from plasmid pRS415a
| Phenotype and conditionsb | pRS415/R+A* | pRS415/R+A+ | pRS415/RΔA* | pRS415/RΔA+ | pRS415 |
|---|---|---|---|---|---|
| Growth (colony diameter, cm) | |||||
| MM-glucose (0.2%) (30°C) | 0.16 ± 0.02 | 0.18 ± 0.03 | 0.15 ± 0.04 | 0.16 ± 0.01 | 0.18 ± 0.02 |
| MM-glucose (0.2%) (42°C) | 0.10 ± 0.02 | 0.21 ± 0.03 | 0.23 ± 0.02 | 0.23 ± 0.05 | 0.25 ± 0.03 |
| MM-succinate (0.1%) | 0c | 0.01 ± 0.02 | 0.09 ± 0.01 | 0.10 ± 0.03 | 0.11 ± 0.02 |
| LB-glucose (0.2%)-amikacin (3 μg/ml) | 0.18 ± 0.04 | 0 | 0 | 0 | 0 |
| LB (pH 9)d | 0.20 ± 0.02 | 0 | 0 | 0 | 0 |
| LB-CuCl2 (4 mM) | 0.21 ± 0.02 | 0 | 0 | 0 | 0 |
| NT-toluidene blue (1.8 mg/ml) | 0.12 ± 0.02 | 0 | 0 | 0 | 0 |
| Growth inhibition (diameter, cm)e | |||||
| LB-H2O2 (30%) | 4.60 ± 0.2 | 3.68 ± 0.12 | 3.63 ± 0.10 | 3.60 ± 0.10 | 3.53 ± 0.07 |
| LB-EDTA (500 mM, pH 8) | 2.00 ± 0.15 | 2.70 ± 0.10 | 2.90 ± 0.10 | 2.80 ± 0.05 | 2.80 ± 0.20 |
| LB-SDS (10%) | 3.1 ± 0.10 | 0 | 0 | 0 | 0 |
| Swarming zone (diameter, cm)f | 2.25 ± 0.05 | 3.20 ± 0.11 | 3.50 ± 0.15 | 3.55 ± 0.15 | 3.52 ± 0.12 |
| Part of population (%) with randomly positioned septumg | 82 ± 4 | 3.00 ± 0.09 | 0 | 2.00 ± 0.02 | 0 |
Ampicillin was used at 100 μg/ml, except in MM-glucose-amikacin and swarming medium (both 35 μg/ml) and MM-succinate (150 μg/ml).
MM, minimal agar medium (pH 7.0) comprising 34 mM NaH2PO4, 66 mM K2HPO4, 20 mM (NH4)2SO4, 1 μM FeSO4, 30 mM MgSO4, 1 mM ZnCl2, 10 μM CaCl2, 0.3 mM isoleucine, 0.3 mM valine, 2 mM thiamine, and ultrapure agarose (17 mg/ml; Seakem LA); LB, Luria-Bertani broth containing Bacto Agar (17 mg/ml; Difco); NT, NaCl (8 mg/ml)-tryptone (10 mg/ml) broth containing Bacto Agar (17 mg/ml; Difco). Growth or growth inhibition was scored after overnight incubation, with the exception of growth on MM-glucose at 30°C (4 days) and MM-succinate (8 days). Growth was scored at 37°C unless otherwise specified.
A zero indicates no growth or no growth inhibition even upon prolonged incubation.
Contains 100 mM CHES (Sigma).
100 μl of a mid-exponential-phase culture was spread on LB in the center of which a paper disk (1.2-cm diameter; Schleicher & Schuell) containing 30 μl of H2O2 or 50 μl of EDTA) or a 5-μl drop of SDS was placed.
Motility was analyzed (overnight) after spotting of 5 μl of a mid-exponential-phase culture at the center of a Difco nutrient (8-mg/ml) soft agar (4-mg/ml) plate supplied with 0.2% glucose and ampicillin.
Cells were grown overnight in 5 ml of LB-ampicillin medium, and growth was scored using phase-contrast microscopy. The percentage of aberrantly dividing cells was calculated from at least 300 cells per culture.