TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristica | Source or reference |
|---|---|---|
| S. paucimobilis | ||
| SYK-6 | Wild type; Nalr Smr | 11 |
| DZ | SYK-6 derivative; desZ::kan; Nalr Smr Kmr | This study |
| DB | SYK-6 derivative; ligB::bla; Nalr Smr Cbr | This study |
| DBZ | SYK-6 derivative; desZ::kan, ligB::bla; Nalr Smr Kmr Cbr | This study |
| P. putida PpY1100 | Nalr Smr | 11 |
| E. coli | ||
| HB101 | supE44 hsdS20 (rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 2 |
| MV1184 | ara Δ(lac-proAB) rspL thi (φ80 lacZΔM15) Δ(srl-recA)306::Tn10 F′ [traD36 proAB+lacIqlacZΔM15] | 41 |
| BL21(DE3) | F−ompT hsdSB(rB− mB−) gal dcm (DE3); T7 RNA polymerase gene under the control of the lacUV5 promoter | 36 |
| Plasmids | ||
| pVK100 | Broad-host-range cosmid vector; Kmr Tcr | 5 |
| pUC19 | Cloning vector; Apr | 43 |
| pBluescript II KS(+) and SK(+) | Cloning vectors; Apr | 33 |
| pT7Blue | Cloning vector; Apr T7 promoter | Novagen |
| pET21a(+) | Expression vector; Apr T7 promoter | Novagen |
| pIK03 | KS(+) with a 1.3-kb EcoRV fragment carrying kan of pUC4K | 19 |
| pK19mobsacB | oriT sacB Kmr | 32 |
| pVK3-1 | pVK100 with approx 20-kb fragment carrying desZ | This study |
| pBX2F | KS(+) with a 7.0-kb XhoI fragment of pVK3-1 | This study |
| pBXSM1 | KS(+) with a 2.7-kb SmaI-XhoI fragment of pBX2F carrying desZ | This study |
| pTDZ | pT7Blue with a 1.0-kb PCR-amplified fragment carrying desZ | This study |
| pEDZA | pET21a(+) with a 1.0-kb NdeI fragment of pTDZ carrying desZ | This study |
| pBDDZ | pBXSMI with an insertion of the 1.3-kb EcoRV fragment carrying kan from pIK03 replacing the 0.8-kb HincII-NruI fragment | This study |
| pKDDZ | pK19mobsacB with a 2.8-kb XbaI-BglII fragment of pBDDZ | This study |
| pSSAB | SK(+) with a 1.5-kb XbaI-SmaI fragment carrying ligAB | This study |
| pKAB | pK19mobsacB with a 1.5-kb XbaI-SmaI fragment of pSSAB | This study |
| pAAB | pKAB with an insertion of the 1.0-kb fragment carrying bla from pUC19 into the MluI site | This study |
a
Abbreviations: Nalr, Smr, Kmr, Cbr, Apr, and Tcr, resistance to nalidixic acid, streptomycin, kanamycin, carbenicillin, ampicillin, and tetracycline, respectively.