TABLE 1.
Bacterial strains and plasmids used
| Strain or plasmid | Relevant characteristicsa | Source or reference |
|---|---|---|
| E. coli | ||
| DH5α | F−recA1 Δ(lacZYA-argF) U169 hsdR17 thi-1 gyra66 supE44 endA1 relA1 Δ(lacZ)M15 | 44 |
| GM2163 | F−ara-14 leuB6 thi-1 fhuA31 lacY1 tsx-78 galK2 galT22 supE44 hisG4 rpsL 136 (Strr) xyl-5 mtl-1 dam13::Tn9 (Camr) dcm-6 mcrB1 hsdR2 mcrA | New England Biolabs |
| HB101 | F−hsdS20 (r−m−) recA13 ara-14 proA2 lacY1 galK2 rpsL20 (Strr) xyl-5 mtl-1 supE44 λ− | 5 |
| P. putida | ||
| 06909 | Ampr, wild type | 59 |
| 06909s21x | Kanr, kanamycin cassette inserted into the LysR family response regulator in wild-type strain 06909 chromosomal locus | This study |
| 06909s22x | Kanr, kanamycin cassette inserted into cadmium-transporting ATPase in wild-type strain 06909 chromosomal locus | This study |
| 06909s23 | Genr, gentamicin cassette inserted into cadR in wild-type strain 06909 chromosomal locus | This study |
| Plasmids | ||
| pUC119 | Apr, cloning vector | 60 |
| pRK415 | Tcr, RK2 derived broad-host-range cloning vector | 21 |
| pRK2013 | Kmr, mobilization helper | 13 |
| pHRP311 | Gmr Smr, an RSF1010 derivative, broad-host-range transcriptional fusion vector | 38 |
| pRKL1 | Tcr, a 3.5-kb HindIII fragment carrying a promoterless lacZ gene from pHRP311 cloned into pRK415 | This study |
| pMGm | Apr Gmr, plasmid carrying a gentamicin resistance gene cassette | 33 |
| pMKm | Apr Kmr, plasmid carrying a kanamycin resistance gene cassette | 33 |
| pRIVS2 | Tcr, pRIV16 carrying a clone expressed on cultured Phytophthora parasitica | 25 |
| pUIVS2 | Apr, pUC119 carrying a 5.5-kb EcoRI fragment from pRIVS2 | 25 |
| pUIVS21 | Apr, pUC119 carrying a 2-kb SphI fragment from pUIVS2 | 25 |
| pUIVS22 | Apr, pUC119 carrying a 1-kb SphI fragment from pUIVS2 with a partial cadmium-transporting ATPase gene | This study |
| pUIVS21K | Apr Kmr, pUIVS21 carrying a 1.8-kb kanamycin cassette from pMKm cloned in a blunt-ended XhoI site of lysR | This study |
| pUIVS22K | Apr Kmr, pUIVS22 carrying a 1.8-kb kanamycin cassette from pMKm cloned in a blunt-ended XhoI site of cadA | This study |
| pRS21k | Tcr Kmr, pRK415 carrying a 3.8-kb SphI fragment from pUIVS21K | This study |
| pRS22k | Tcr Kmr, pRK415 carrying a 2.8-kb SphI fragment from pUIVS22K | This study |
| pRS23s | Tcr Gmr, pRCD12 carrying a 1.9-kb PstI fragment blunt-ended gentamicin cassette from pMGm cloned in a blunt-ended ClaI site of cadR | This study |
| pRCD12 | Tcr, a 4.2-kb PstI fragment carrying cadmium resistance genes in pRK415 | This study |
| pRCD22 | Tcr, the 4.2-kb PstI fragment in pRCD12 is reverse oriented against lac promoter in pRK415 | This study |
| pRCD13 | Tcr, a 3.0-kb PstI-HindIII fragment of pRCD12 cloned in pRK415 | This study |
| pRCD14 | Tcr, a 1.0-kb PstI-XhoI fragment was removed from pRCD13 | This study |
| pUCD12 | Apr, a 3.0-kb PstI-HindIII fragment of pRCD12 in pUC119 | This study |
| pUCD30 | Apr, pUC129 carrying a 2-kb ClaI-XhoI fragment of pRCD12 | This study |
| pRCD31 | Tcr, pRKL1 carrying cadA promoter region, an 800-bp EcoRI-KpnI fragment of pUCD30 | This study |
| pRCD32 | Tcr, pRKL1 carrying cadR promoter region, an 800-bp PstI-KpnI fragment of pUCD30 | This study |
a
Ampr, chromosomal ampicillin resistance; Kanr, chromosomal kanamycin resistance; Genr, chromosomal gentamicin resistance; Apr, ampicillin resistance; Tcr, tetracycline resistance; Kmr, kanamycin resistance; Gmr, gentamicin resistance; Smr, streptomycin resistance.