TABLE 1.
Microorganisms and plasmids used
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | F−recA1 Δ(lacZYA-argF)U169 hsdR17 thi-1 gyrA66 supE44 endA1 relA1 φ80dlac Δ(lacZ)M15 | 40 |
| HB101 | F−hsdA20 (r− m−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 xyl-5 mtl-1 supE44 λ− | 2 |
| P. putida strains | ||
| 06909 | Wild type; Ampr | 54 |
| 06909u1 | pyrB; uracil auxotroph; Ampr Kanr | This study |
| 06909u2 | pyrB; uracil auxotroph; Ampr Kanr | This study |
| P. parasitica M191 | 54 | |
| Plasmids | ||
| pUC119 | Apr; cloning vector | 55 |
| pRK415 | Tcr; RK2-derived broad-host-range cloning vector | 20 |
| pHRP309 | Gmr; RSF1010-based broad-host-range transcriptional fusion vector | 34 |
| pHSK728 | Apr Smr; Tn7 delivery plasmid | 44 |
| pHSK828 | Apr Smr; pHSK728 derivative; a lux operon in pHSK728 was replaced with an ice nucleation gene | Unpublished data |
| pDSK509 | Kmr; RSF1010-derived broad-host-range cloning vector | 20 |
| pRK2013 | Kmr; mobilization helper | 10 |
| pDAC41 | Apr; pyrB cloned in pUC119 at HincII site | This study |
| pDAC42 | Apr Kmr; 1.2-kb kanamycin cassette inserted in pDAC41 | This study |
| pDAC43 | Apr; pyrBC′ cloned in pUC119 at HincII site | This study |
| pDAC54 | Apr; Φ(pyrBC′-lacZ) cloned in pHSK828 | This study |
| pRIV11 | Tcr; pRK415 carrying a 1.8-kb SphI-EcoRI fragment (blunt ended) from pDAC42 containing the pyrB::km fragment | This study |
| pRIV13 | Tcr; pRK415 carrying a 2.4-kb HindIII-BamHI fragment from pDAC43 containing pyrBC′ | This study |
| pRIV16 | Tcr; pRK415 carrying a 6.1-kb HindIII-BamHI fragment from pDAC54 containing a promoterless fusion; Φ(pyrBC′-lacZ) in opposite orientation compared to lac promoter | This study |
| pRIV16C | Tcr; pRK415 carrying a promoterless fusion; Φ(pyrBC′-lacZ) in same orientation as lac promoter | This study |
| pRIVA1 | Tcr; pRIV16 carrying a constitutive promoter | This study |
| pRIVS1 | Tcr; pRIV16 carrying a clone expressed on fungal plates | This study |
| pRIVS2 | Tcr; pRIV16 carrying a clone expressed on fungal plates | This study |
| pRIVS3 | Tcr; pRIV16 carrying a clone expressed on fungal plates | This study |
| pRIVS4 | Tcr; pRIV16 carrying a clone expressed on fungal plates | This study |
| pRIVS5 | Tcr; pRIV16 carrying a clone expressed on fungal plates | This study |
| pRIVS13 | Tcr; 3.9-kb EcoRI fragment from pRIVS1 transferred into pRIV16 | This study |
| pRIVS20 | Tcr; pRIV16 carrying a 1.4-kb PstI fragment from pRIVS2; the fragment has the same orientation as Φ(pyrBC′-lacZ) of pRIV16 | This study |
| pRIVS26 | Tcr; pRIV16 carrying a 1.4-kb PstI fragment from pRIVS2; the fragment has the opposite orientation compared to Φ(pyrBC′-lacZ) of pRIV16 | This study |
| pUIVS1 | Apr; pUC119 carrying a 5.4-kb KpnI fragment from pRIVS1 | This study |
| pUIVS11 | Apr; pUC119 carrying a 1.4-kb EcoRI-HindIII fragment from pUIVS1 | This study |
| pUIVS2 | Apr; pUC119 carrying a 5.5-kb EcoRI fragment from pRIVS2 | This study |
| pUIVS21 | Apr; pUC119 carrying a 2-kb SphI fragment from pUIVS2 | This study |
| pUIVS3 | Apr; pUC119 carrying a 4.2-kb EcoRI fragment from pRIVS5 | This study |
| pUIVS4 | Apr; pUC119 carrying a 2-kb EcoRI-SphI fragment from pRIVS4 | This study |
| pUIVS5 | Apr; pUC119 carrying a 4.5-kb EcoRI fragment from pRIVS5 | This study |
a
Ampr, chromosomal ampicillin resistance; Kanr, chromosomal kanamycin resistance; Apr, ampicillin resistance; Tcr, tetracycline resistance; Smr, streptomycin resistance; Gmr, gentamicin resistance; Kmr, kanamycin resistance.