TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Properties or genotypea | Source or reference |
|---|---|---|
| P. aeruginosa | ||
| K767 | PAO1, wild type | 47 |
| K1491 | K767 ΔmexR | 75 |
| K1119 | K767 ΔmexAB-oprM | 41 |
| K870 | Smr derivative of K767 | 65 |
| K2278 | K870 ΔmexA | This study |
| K2274 | K870 ΔmexRA | This study |
| K2275 | K767 ΔmexRAB | This study |
| E. coli | ||
| DH5α | endA hsdR17 supE44 thi-1 recA1 gyrA relA1 Δ(lacZYA-argF)U169 deoR [φ80 dlacΔ(lacZ)M15] | 3 |
| S17-1 | thi pro hsdR recA Tra+ | 71 |
| SU101 | lexA71::Tn5 (Def)sulA211 sulA::lacZ | 13 |
| Δ(lacIPOZYA)169/F′ lacIqlacZΔM15::Tn9 | ||
| Plasmids | ||
| pAK1900 | E. coli-P. aeruginosa shuttle cloning vector; MCS; Cbr/Apr | A. Kropinski, Queens University |
| pPV2 | pAK1900 carrying mexR-mexAB-oprM and flanking DNA on a 9-kb XhoI fragment | 62 |
| pPV6 | pAK1900 carrying mexR-mexAB-oprM and flanking DNA on a 5.5-kb XhoI-HindIII fragment | 62 |
| pRSP48 | pAK1900::mexA | This study |
| pEX18Tc | Gene-replacement vector; sacB Tcr | 23 |
| pDN1 | pEX18Tc:: ΔmexRA | This study |
| pDN2 | pEX18Tc:: ΔmexRAB | This study |
| pRK415 | Broad-host-range cloning vector; Tcr | 32 |
| pRSP17 | pRK415::mexAB-oprM | 73 |
| pDN3 | pRK415::mexA (WT) | This study |
| pDN4 | pRK415::mexA* (P68S) | This study |
| pDN5 | pRK415::mexA* (G72S) | This study |
| pDN6 | pRK415::mexA* (A108T) | This study |
| pDN7 | pRK415::mexA* (V129M) | This study |
| pDN8 | pRK415::mexA* (T256I) | This study |
| pDN9 | pRK415::mexA* (V264E) | This study |
| pDN10 | pRK415::mexA* (N270S) | This study |
| pDN11 | pRK415::mexA* (H279Y) | This study |
| pDN12 | pRK415::mexA* (V286A) | This study |
| pDN13 | pRK415::mexA* (G297D) | This study |
| pDN26 | pRK415::mexA* (L91P) | This study |
| pDN27 | pRK415::mexA* (L110P) | This study |
| pDN28 | pRK415::mexA* (N270S, S251P) | This study |
| pDN29 | pRK415::mexA* (G297D, L155P) | This study |
| pMS604 | pUC-like vector carrying a LexA1-87 WT-Fos zipper fusion and a ColE1 replicon; Tcr | 66 |
| pDN14 | pMS604::mexA (WT) | This study |
| pDN15 | pMS604::mexA* (P68S) | This study |
| pDN16 | pMS604::mexA* (G72S) | This study |
| pDN17 | pMS604::mexA* (A108T) | This study |
| pDN18 | pMS604::mexA* (V129M) | This study |
| pDN19 | pMS604::mexA* (T256I) | This study |
| pDN20 | pMS604::mexA* (V264E) | This study |
| pDN21 | pMS604::mexA* (N270S) | This study |
| pDN22 | pMS604::mexA* (H279Y) | This study |
| pDN23 | pMS604::mexA* (V286A) | This study |
| pDN24 | pMS604::mexA* (G297D) | This study |
| pDN28 | pMS604::mexA* (L91P) | This study |
| pDN29 | pMS604::mexA* (L110P) | This study |
| pVLT31 | Broad-host-range, low-copy-number cloning vector lacIq Tcr | 11 |
| pRE38 | pVLT31::mexB-6his | This study |
| pMMB206 | Broad-host-range, low-copy-number cloning vector; lacIq Cmr | 51 |
| pDN25 | pMMB206::mexB-6his | This study |
| pET21d(+) | His tag vector used to generate His6 C-terminal fusions to products of cloned genes; Apr | Novogen |
| pXZL05 | pET21d(+)::mexA | This study |
a
Smr, streptomycin resistance; Cbr, carbenicillin resistance; Tcr, tetracycline resistance; Apr, ampicillin resistance; Cmr; chlorampheicol resistance; mexB-6his, a mexB gene engineered to produce a MexB protein with six C-terminal histidine residues; MCS, multiple cloning site. Mutations in the MexA proteins encoded by the plasmids are shown in parentheses WT, wild-type MexA.