TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristicsa | Reference or source |
|---|---|---|
| Pseudomonas | ||
| P. alcaligenes NCIB 9867 | Wild type, 2,5-xylenol+ 3,5-xylenol+m-cresol+ | 16 |
| P. alcaligenes P25X14 | Strr/Spcr, pRA14 | This study |
| P. alcaligenesc P25X19 | Strr/Spcr, pRA19 | This study |
| P. alcaligenes P25X20 | Kanr, Tn5 insertion into NCIB 9867 | This study |
| P. putida RA713 | Plasmid-free derivative of NCIB 9869 | 17 |
| P. putida RA713RifNal | Rifr Nalr, spontaneous mutant of RA713 | This study |
| P. putida KT2440 | Restriction-deficient derivative of P. putida mt-2 | 2 |
| P. putida KT2440RifNal | Rifr Nalr, spontaneous mutant of KT2440 | This study |
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | 32 |
| DH10B | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 Δ(lac)X74 deoR recA1 araD139 Δ(ara leu)7697 galU galK rpsL endA1 nupG | 25 |
| DF1 | F mcrB mrr hsdS20(rB− mB−) supE44 ara-14 galK2 lacY1 proA2 rpsL20 (Strr) xyl-5 λ− leuB6 mtl-1 Δ(srl-recA)306 γδ−tnpA+ Camr | 43 |
| S17-1 | thi pro hsdR hsdM+ recA RP4 tra | 36 |
| Plasmids | ||
| pRA2 | Cryptic endogenous plasmid from P. alcaligenes NCIB 9867 | 23 |
| pVK182 | pRA2 minireplicon: pRA2 replication region cloned into BamHI and gene cassette from Tn5 conferring KAN resistance cloned into the HindIII sites of pUC18 | 23 |
| pHP45Ω | Source of ΩStrr/Spcr | 30 |
| pSUP202 | pBR325 derivative, Ampr Camr Tetrmob+ | 36 |
| pRA14 | pRA2 pac25I.R::ΩStrr/Spcr (replaced nt 20373 to 20396) | This study |
| pRA19 | pRA2 Tn5563::ΩStrr/Spcr (inserted at nt 13355) | This study |
| pRK415 | IncP TetrlacZ mob, broad-host-range cloning vector | 21 |
| pPSK1 | 19,386-bp PstI fragment from pRA2 cloned into pRK415 | This study |
| pPSK2 | 6,311-bp PstI fragment from pRA2 cloned into pRK415 | This study |
| pPSK3 | 3,405-bp PstI fragment from pRA2 cloned into pRK415 | This study |
| pPSK4 | 2,794-bp PstI fragment from pRA2 cloned into pRK415 | This study |
| pPSK5 | 847-bp PstI fragment from pRA2 cloned into pRK415 | This study |
| pDelta2 | pBR322 replicon, Ampr Kanr Sucs StrslacZ | 43 |
| pVLT33 | IncQ KanrPtac lacIqmob, broad-host-range cloning vector | 10 |
| pMMB67EH | IncQ AmprPtac lacIqmob, broad-host-range cloning vector | 12 |
| RK2/RP4 | IncPα Ampr Kanr Tetrtra, endogenous plasmid | 9 |
a
Ampr, Kanr, Strr, Spcr, Tetr, Camr, Rifr, and Nalr indicate resistance to the respective antibiotics. Strs and Sucs indicate sensitivity to STR and sucrose, respectively. mob, mobilized by RK2/RP4 transfer functions. tra, RK2/RP4 transfer functions. nt, nucleotide.