TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Relevant characteristic(s)a | Reference or source |
|---|---|---|
| S. paucimobilis strains | ||
| SYK-6 | Wild type; Nalr Smr | 29 |
| DW | SYK-6 derivative, ligW::kan Nalr Smr Kmr | This study |
| DW2 | SYK-6 derivative, ligW2::kan Nalr Smr Kmr | This study |
| DDW | SYK-6 derivative, ligW::kan ligW2::bla Nalr Smr Kmr Cbr | This study |
| Dorf1 | SYK-6 derivative, orf1::kan Nalr Smr Kmr | This study |
| Dorf3 | SYK-6 derivative, orf3::kan Nalr Smr Kmr | This study |
| P. putida PpY101 | Nalr Smr | 17 |
| E. coli strains | ||
| JM109 | recA1 supE44 endA1 hsdR17 gyrA96 relA1 thiΔ(lac-proAB) F′[traD36 proAB+lacIqlacZΔM15] | 56 |
| S17-1 | recA; harbors the tra genes of plasmid RP4 in the chromosome, proA thi | 51 |
| HB101 | supE44 hsdS20 (rB− mB−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 | 5 |
| Plasmids | ||
| pUC18 | Cloning vector; Apr | 56 |
| pBluescript II KS(+) | Cloning vector; Apr | 50 |
| pVK100 | Broad-host-range cosmid vector, Kmr Tetr | 14 |
| pRK2013 | Kmr Tra+ Mob+ | 16 |
| pDEC6 | pVK100 with an approximately 20-kb fragment carrying ligW2 | This study |
| pU351 | pUC18 with a 3.6-kb SalI fragment carrying ligW2 | This study |
| pU352 | pUC18 carrying the same fragment as pU351 in the opposite direction | This study |
| pA1, pA5, pA9, pA13, pA14 | Deletion derivatives of pU351 | This study |
| pB15, pB16, pB19, pB26, pB28, pB32 | Deletion derivatives of pU352 | This study |
| pW2 | KS(+) with a 1.1-kb SmaI-SacII fragment carrying ligW2 | This study |
| pVKW2 | pVK100 carrying the same fragment as pW2 | This study |
| pVK7E2 | pVK100 with a 7.0-kb EcoRI fragment carrying ligW | 41 |
| pUC4K | Apr/Kmr | 53 |
| pK19mobsacB | oriT sacB Kmr | 49 |
| pU351Km | pU351 with insertion of kan from pUC4K into an EcoRV site | This study |
| pDW2Km | pK19mobsacB with a 4.8-kb SalI fragment from pU351Km | This study |
| pK19U351 | pK19mobsacB with a 3.6-kb SalI fragment from pU351 | This study |
| pDW2Ap | pK19U351 with insertion of bla from pUC18 into an EcoRV site | This study |
| pKS7E2 | KS(+) with a 7.0-kb EcoRI fragment of pVK7E2 | 41 |
| pKV44 | Deletion derivative of pKS7E2 carrying ligW | 41 |
| pKV44Km | pKV44 with insertion of kan from pUC4K into a SalI site | This study |
| pDW | pK19mobsacB with a 3.6-kb SacI-XbaI fragment from pKV44Km | This study |
| pUC11 | pUC19 with a 2.8-kb BamHI fragment carrying orf1 | This study |
| pUC11Km | pUC11 with insertion of kan from pUC4K into a SalI site | This study |
| pDorf1 | pK19mobsacB with a 4.0-kb BamHI fragment from pUC11Km | This study |
| pSV322 | KS(+) with a 2.8-kb EcoRV-SalI fragment carrying orf3 | This study |
| pSV322Km | pSV322 with insertion of kan from pUC4K into a HindIII site | This study |
| pDorf3 | pK19mobsacB with a 4.0-kb EcoRI-SalI fragment from pSV322Km | This study |
| pHN139F | pUC18 with a 10.5-kb EcoRI fragment carrying ligJAB, part of ligC, ligK, and ligI | 35 |
| pDE20 | pVK100 with an approximately 20-kb EcoRI fragment carrying part of ligM, metF, and ligH | 37 |
a
Abbreviations: Nalr, Smr, Kmr, Cbr, Tetr and Apr, resistance to nalidixic acid, streptomycin, kanamycin, carbenicillin, tetracycline, and ampicillin, respectively.