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. 2014 Dec;196(23):4152–4162. doi: 10.1128/JB.02202-14

FIG 3.

FIG 3

Oxygen-inhibited and NarK2-dependent reduction of nitrate by the mycelium of S. coelicolor. (A) Nitrite production by the mycelium of S. coelicolor strain NM59 (Δnar-1 Δnar-3) after 2.5 h of incubation in MOPS buffer in the presence of 10 mM nitrate was monitored. Mycelium was incubated aerobically, anaerobically, or in standing liquid culture. One of each variant was additionally incubated with 200 μg/ml chloramphenicol (CLM). (B) Relative (rel.) nitrite release of mutant strains of S. coelicolor A3(2) in comparison to that of wild-type strain M145 (*, 100% relative nitrite release is equivalent to approximately 2 mM nitrite). Nitrite was determined after 2.5 h of incubation in MOPS buffer with 10 mM NO3. The incubation was carried out with standing liquid cultures. (C) Western blot analysis with anti-NarG2 peptide antibodies of extracts (60 μg) of mycelium derived from the indicated mutants. After separation in SDS-polyacrylamide gels (7.5% [wt/vol] polyacrylamide), the samples were probed with polyclonal peptide antibodies specific for NarG2. The migration position of NarG2 (139 kDa) is shown on the right. An unidentified cross-reacting band migrating at approximately 70 kDa acted as a loading control. (D) Schematic representation of the genetic locus upstream of the narG2H2J2I2 operon (SCO0216 to SCO0219) in the S. coelicolor A3(2) genome.