TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| Sphingobium sp. strains | ||
| SYK-6 | Wild type; Nalr Smr | 68 |
| SME110 | SYK-6 derivative; phcC::kan; Nalr Smr Kmr | This study |
| SME111 | SYK-6 derivative; phcD::kan; Nalr Smr Kmr | This study |
| SME112 | SYK-6 derivative; phcC-phcD::kan; Nalr Smr Kmr | This study |
| E. coli strains | ||
| BL21(DE3) | F− ompT hsdSB(rB− mB−) gal dcm (DE3); T7 RNA polymerase gene under the control of the lacUV5 promoter | 69 |
| HB101 | recA13 supE44 hsd20 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 70 |
| NEB 10-beta | araD139 Δ(ara-leu)7697 fhuA lacX74 galK (ϕ80 ΔlacZ M15) recA1 endA1 nupG rpsL (Smr) Δ(mrr-hsdRMS-mcrBC) | New England Biolabs |
| Plasmids | ||
| pT7Blue | Cloning vector; Apr | Novagen |
| pBluescript II KS(+) | Cloning vector; Apr | 71 |
| pET-16b | Expression vector; T7 promoter; Apr | Novagen |
| pJB866 | RK2 broad-host-range expression vector; Tcr Pm xylS | 72 |
| pJB861 | RK2 broad-host-range expression vector; Kmr Pm xylS | 72 |
| pK19mobsacB | oriT sacB; Kmr | 73 |
| pIK03 | pBluescript II KS(+) with a 1.3-kb EcoRV fragment carrying kan of pUC4K; Apr Kmr | 28 |
| pKS09480-500 | pBluescript II KS(+) with a 3.9-kb PCR-amplicon carrying phcC and phcD | This study |
| pKS09480 | pBluescript II KS(+) with a 2.4-kb BglII-BamHI fragment carrying phcC from pKS09480-500 | This study |
| pKS09500 | pBluescript II KS(+) with a 2.7-kb XhoI-SacI fragment carrying phcD from pKS09480-500 | This study |
| pKS09480K | pKS09480 with a 1.3-kb EcoRV fragment carrying kan from pIK03 into HincII site of phcC | This study |
| pKS09500K | pKS09500 with a 1.3-kb EcoRV fragment carrying kan from pIK03 into HincII site of phcD | This study |
| pKmb09480K | pK19mobsacB with a 3.7-kb KpnI-BamHI fragment carrying disrupted phcC of pKS09480K | This study |
| pKmb09500K | pK19mobsacB with a 4.0-kb KpnI-SacI fragment carrying disrupted phcD of pKS09500K | This study |
| pT09480-500 | pT7Blue with a 3.9-kb PCR-amplicon carrying phcC and phcD | This study |
| pT09480-500K | pT09480-500 with a 1.3-kb EcoRV fragment carrying kan from pIK03 into HincII site of phcC and phcD | This study |
| pKmb09480-500K | pK19mobsacB with a 3.0-kb SphI-BamHI fragment carrying disrupted phcC and phcD of pT09480-500K | This study |
| pT09480 | pT7Blue with a 1.7-kb PCR-amplicon carrying phcC | This study |
| pT09500 | pT7Blue with a 1.8-kb PCR-amplicon carrying phcD | This study |
| pT09480Xba | pT7Blue with a 1.8-kb XbaI-BamHI fragment carrying phcC from pT09480 | This study |
| pT09500Xba | pT7Blue with a 1.9-kb XbaI-BamHI fragment carrying phcD from pT09500 | This study |
| pET09480 | pET-16b with a 1.7-kb NdeI-BamHI fragment carrying phcC from pT09480 | This study |
| pET09500 | pET-16b with a 1.8-kb NdeI-BamHI fragment carrying phcD from pT09500 | This study |
| pJBVI09480 | pJB866 with a 1.8-kb HindIII-BamHI fragment carrying phcC from pT09480Xba | This study |
| pJBVI09500 | pJB866 with a 1.9-kb HindIII-BamHI fragment carrying phcD from pT09500Xba | This study |
| pJBI09480 | pJB861 with a 1.8-kb KpnI-BamHI fragment carrying phcC from pJBVI09480 | This study |
| pJBI09500 | pJB861 with a 1.9-kb KpnI-BamHI fragment carrying phcD from pJBVI09500 | This study |
a
Kmr, Nalr, Smr, Apr, and Tcr, resistance to kanamycin, nalidixic acid, streptomycin, ampicillin, and tetracycline, respectively.