Table 1.
Strains and plasmids used in this study
| Strain or plasmid | Characteristics | Source or reference(s) |
|---|---|---|
| Strains | ||
| Pseudomonas | ||
| Pseudomonas sp. HZN6 | Apr; Wild type, nicotine degrader; G− | 9, 10 |
| P. putida KT2440 | Apr; non-nicotine-degrading strain | 25 |
| N6Δnox | Apr Kmr; nox::Kmr mutant of HZN6 | This study |
| N6ΔnoxC | Apr Kmr Gmr; N6Δnox containing pBB-nox | This study |
| KT-nox | Apr Gmr; KT2440 containing pBB-nox | This study |
| E. coli | ||
| DH5α | λ− ϕ80dlacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK−) supE44 thi-1 gyrA relA1 | TaKaRa |
| HB101(pRK2013) | Conjugation helper strain | Laboratory stock |
| SM10λpir | thi thr leu tonA lacY supE recA::RP-4-Tc::Mu (λ pir) | Laboratory stock |
| DH-nox | Gmr; DH5α containing pBB-nox | This study |
| Plasmids | ||
| pMD18-T | Apr; T-A clone vector | TaKaRa |
| pJQ200SK | Gmr mob+ orip15A lacZα+ sacB; suicide vector | 29 |
| pBBR1-MCS5 | Gmr; broad-host-range cloning vector | 27 |
| pJQΔnox | Gmr; ApaI-SacI fragment containing nox inserted into pJQ200SK where nox was disrupted by kanamycin resistance gene | This study |
| pBB-orf12nox | Gmr; KpnI-SacI fragment containing orf1, orf2, and nox inserted into pBBR1-MCS5 | This study |
| pBB-orf2nox | Gmr; KpnI-SacI fragment containing orf2 and nox inserted into pBBR1-MCS5 | This study |
| pBB-nox | Gmr; KpnI-SacI fragment containing nox inserted into pBBR1-MCS5 | This study |
| pBB-orf12noxH456R | Gmr; pBB-orf12nox with a mutation of His456 to Arg | This study |
| pBB-orf2noxH456R, | Gmr; pBB-orf2nox with a mutation of His456 to Arg | This study |
| pBB-noxH456R, | Gmr; pBB-nox with a mutation of His456 to Arg | This study |
| pBB-Δmotif | Gmr; pBB-nox with deletion of FAD-binding motif GxGxxG | This study |