TABLE 2.
Bacterial strains and plasmids used or constructed in this study
| Strain or plasmid | Genotype or relevant characteristicsa | Reference or source |
|---|---|---|
| Strains | ||
| E. coli strains | ||
| Stellar | E. coli HST08 strain for cloning of plasmids | Clontech |
| JM109 | E. coli strain for cloning of plasmids | Promega |
| P. aeruginosa strains | ||
| PA14 | Wild type | 68 |
| CW26 | ponATP::Gm | This study |
| CW28 | mrcBTP::Gm | This study |
| CW29 | ΔponATP | This study |
| CW30 | ΔponATP pbpA::Gm | This study |
| CW31 | ΔmrcBTP | This study |
| CW34 | ΔmrcBTP pbpA::Gm | This study |
| CW36 | pbpA::Gm | This study |
| CW37 | ΔpbpA | This study |
| CW46 | ΔftsI attB::PBAD-gfp-ftsI | This study |
| CW47 | ΔponATP ΔmrcBTP attB::PBAD-gfp-ponA | This study |
| CW49 | ΔponAGT attB::PBAD-gfp-ponA | This study |
| CW50 | ΔponAGT ΔmrcBGT attB::PBAD-gfp-ponA | This study |
| CW52 | ΔponATP attB::PBAD-gfp-ponA | This study |
| CW57 | PA14 containing pMP7605-PBP1a | This study |
| CW58 | PA14 containing pMP7605-PBP1b | This study |
| CW59 | PA14 containing pMP7605-PBP2 | This study |
| CW60 | PA14 containing pMP7605-PBP3 | This study |
| CW61 | PA14 containing pJN-flhF-mCherry | This study |
| CW80 | pbpC::Gm | This study |
| CW81 | ΔftsI pbpC::Gm attB::PBAD-gfp-ftsI | This study |
| Plasmids | ||
| pEX18ApGw | Suicide vector for gene knockout in P. aeruginosa, Gmr CBCr | 62 |
| pEX18ApGW-pbp1aTP | Suicide vector for deletion of the PBP1a TPase domain, Gmr CBCr | This study |
| pEX18ApGW-pbp1bTP | Suicide vector for deletion of the PBP1b TPase domain, Gmr CBCr | This study |
| pEX18ApGW-pbp2 | Suicide vector for deletion of pbp2, Gmr CBCr | This study |
| pEX18ApGW-pbp3 | Suicide vector for deletion of pbp3, Gmr CBCr | This study |
| pEX18ApGW-pbp3x | Suicide vector for deletion of pbp3x, Gmr CBCr | This study |
| pEX18ApGW-pbp1aGT | Suicide vector for deletion of PBP1a GTase domain, Gmr CBCr | This study |
| pEX18ApGW-pbp1bGT | Suicide vector for deletion of PBP1b GTase domain, Gmr CBCr | This study |
| pMini-CTX1 | Integration vector in P. aeruginosa, Tcr | 69 |
| pBAD24 | Plasmid with PBAD promoter induced by l-arabinose, CBCr | 35 |
| pCAB | Derivative of pMini-CTX1 containing PBAD promoter, Tcr | This study |
| pCAB-gfp-pbp1a | Derivative of pCAB containing PBAD-gfp-ponA, Tcr | This study |
| pCAB-gfp-pbp3 | Derivative of pCAB containing PBAD-gfp-ftsI, Tcr | This study |
| pMP7605 | E. coli-P. aeruginosa shuttle vector expressing mCherry, Gmr | 61 |
| pMP7605-PBP1a | Derivative of pMP7605 expressing mCherry-PBP1a, Gmr | This study |
| pMP7605-PBP1b | Derivative of pMP7605 expressing mCherry-PBP1b, Gmr | This study |
| pMP7605-PBP2 | Derivative of pMP7605 expressing mCherry-PBP2, Gmr | This study |
| pMP7605-PBP3 | Derivative of pMP7605 expressing mCherry-PBP3, Gmr | This study |
| pJN-flhF-mCherry | Derivative of pJN105 expressing FlhF-mCherry, Gmr | 46 |
a
CBCr, carbenicillin resistant; Gmr, gentamicin resistant; Tcr, tetracycline resistant.