TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or origin |
|---|---|---|
| Strains | ||
| R. opacus | ||
| SAO101 | 4-NP degrader, wild type, 4-NP+ | 12 |
| SDC1 | npcC gene-disrupted mutant of SAO101, 4-NP− (Kmr) | This study |
| SDA1 | npcA gene-disrupted mutant of SAO101, 4-NP− (Kmr) | This study |
| E. coli | ||
| JM109 | recA1 endA1 gyrA96 thi hsdR17 (rK− mK+) e14− (mcrA) supE44 relA1 Δ(lac-proAB)/F′[traD36 proAB+lacIqlacZΔM15] | Takara Shuzo |
| Rosetta(DE3)pLysS | F−ompT hsdSβ (rB− mB−) gal dcm, lacY1 (DE3) pLysSRARE (Cmr) | Novagen |
| Plasmids | ||
| pBluescriptII KS, SK | Cloning vector, Apr | Stratagene |
| pBS-aphII | Cloning vector, Apr Kmr | This study |
| pHQ157 | Strain SAO101 cosmid library clone containing npc genes, Apr | This study |
| pET17b | Expression vector, Apr | Novagen |
| pETnpcBA | pET17b derivative carrying the npcBA genes | This study |
| pETnpcB | pET17b derivative carrying the npcB gene | This study |
| pETnpcA | pET17b derivative carrying the npcA gene | This study |
| pETnpcC | pET17b derivative carrying the npcC gene | This study |
a
4-NP+, growth on 4-NP; 4-NP−, no growth on 4-NP; Apr, ampicillin resistance; Cmr, chloramphenicol resistance; Kmr, kanamycin resistance.