Table 2.
HAP- and chlorate-sensitivity of wild-type, moa, and mogA strains.
| Strain | Relative genotype | HAP sensitivitya | Chlorate sensitivityb KClO3 (mM) | ||
|---|---|---|---|---|---|
| 15 | 5 | 0.5 | |||
| KP7600 derivatives: | |||||
| KP7600 | wild-type | R | S | R | R |
| JD26193 | moaA::mini-Tn10kan | S | R | R | R |
| JD26196 | moaC::mini-Tn10kan | S | R | R | R |
| JD26197 | moaD::mini-Tn10kan | S | R | R | R |
| JD26198 | moaE::mini-Tn10kan | S | R | R | R |
| JD26194 | moaB::mini-Tn10kan | R | R | R | R |
| JD26195 | moaB::mini-Tn10kan | R | R | R | R |
| KP7600-ΔB | ΔmoaB (in-frame) | R | S | R | R |
| NR10836 derivatives: | |||||
| NR10836 | wild-type | R | S | S | I |
| NR17378 | ΔmoaA (in-frame) | S | R | R | R |
| NR17380 | ΔmoaC (in-frame) | S | R | R | R |
| NR17381 | ΔmoaD (in-frame) | S | R | R | R |
| NR17382 | ΔmoaE (in-frame) | S | R | R | R |
| NR17427 | moaB::mini-Tn10kan | R | S | R | R |
| NR17428 | moaB::mini-Tn10kan | R | S | R | R |
| NR17379 | ΔmoaB (in-frame) | R | S | S | I |
| NR17383 | ΔmogA::kan | S | R | R | R |
| NR17383-p[moaB]c | ΔmogA:: kan, p[moaB]c | S | R | R | R |
| NR10836-p[moaB]c | wt, p[moaB]c | R | S | S | I |
The sensitivity to HAP-induced killing was tested in spot-tests as described in Materials and methods. “R” indicates HAP-resistant phenotype (as “wild-type” in Fig. 1C), “S” indicates HAP-sensitive phenotype (as ΔmoaA,C,D,E in Fig. 1C).
Cells were plated on LB-chlorate plates and incubated 48 h at 37° C under anaerobic conditions (see Materials and methods). Under these conditions, chlorate-sensitive strains do not form colonies (chlorate-sensitive phenotype, “S”) or form very small colonies (chlorate-inhibited phenotype, “I”), whereas chlorate-resistant strains (chlorate-resistant phenotype, “R”) form colonies of the same size as on regular LB plates.
p[moaB] indicates vector pBluescript II SK(+) containing the moaB gene (see Materials and methods). Identical results were obtained with strain NR17391 carrying plasmid pET3a-moaB (data not shown). All tests with plasmid-containing strains were performed in the presence of ampicillin and 0.5 mM IPTG.