TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Descriptiona | Reference and/or source |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| DH5α | F− ϕ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 λ− thi-1 gyrA96 relA1 | Invitrogen |
| JK10 | clpP::cat Δlon slyD::kan λDE3 | 45 |
| ST18 | S17 λpir ΔhemA | 46 |
| ER2556(pLysS) | F− λ− fhuA2 lon ompT lacZ::T7 gene I gal sulA11 Δ(mcrC-mrr)114::IS10 R(mcr-73::mini-Tn10) | 47 |
| Pseudomonas aeruginosa | ||
| PAO1 | University of Washington Collection, 48 | |
| ΔP1 mutant | ΔClpP1 (ΔPA1801) | This study |
| ΔP2 mutant | ΔClpP2 (ΔPA3326) | This study |
| ΔP1P2 mutant | ΔClpP1 ΔClpP2 (ΔPA1801 ΔPA3326) | This study |
| ΔfliC mutant | PW2970 transposon mutant | 48 |
| ΔpilA mutant | PW8622 transposon mutant | 48 |
| Saccharomyces cerevisiae InvSc1 | MATa his3D1 leu2 trp1-289 ura3-52 MAT his3D1 leu2 trp1-289 ura3-52 | Life Technologies |
| Plasmids | ||
| pET23b | Apr, expression plasmid | Novagen |
| pET28b | Kmr, expression plasmid | Novagen |
| pMQ30 | Gmr, tool for gene deletion | 18 |
| ClpP1-pMQ30 | Gene deletion tool | This study |
| ClpP2-pMQ30 | Gene deletion tool | This study |
| pMQ71 | Apr Gmr, shuttle expression plasmid | 18 |
| ClpP1-pMQ71 | ClpP1 complementation plasmid | This study |
| ClpP2-pMQ71 | ClpP2 complementation plasmid | This study |
| pET23b.smt3 | Polyhistidine-tagged Smt3 construct | 49 |
| pBH111-pET23b | ClpP1 with C-terminal polyhistidine tag | This study |
| pBH213-pET23b | ClpP2 with C-terminal Strep II tag | This study |
| pBH304-pET28b | ClpX with N-terminal thrombin-cleavable polyhistidine tag | This study |
| pBH402-pET23b | ClpA with N-terminal Ulp1-cleavable polyhistidine tag and Smt3 SUMO tag | This study |
a
R, rearrangement; Apr, ampicillin resistance; Kmr, kanamycin resistance; Gmr, gentamicin resistance.