TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or relevant characteristics | Reference or Source |
|---|---|---|
| Strains | ||
| P. aeruginosa | ||
| PAO1 | Wild type, serotype O5 | 25 |
| PAK | Wild type, serotype O6; background for IVET | 62 |
| PAK-AR2 | PAK lacking the purEK gene; Spr Smr | 62 |
| 9.27A | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 9.27B | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 9.27C | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 9.27D | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 9.27E | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 9.29 | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| 3.15 | Biofilm-induced IVET clone; Tcr Pipr Spr Smr | This study |
| PA0240::Gmr | PAO1 with insertional mutation in PA0240 | This study |
| PA3710::Gmr | PAO1 with insertional mutation in PA3710 | This study |
| PA3782::Gmr | PAO1 with insertional mutation in PA3782 | This study |
| PA5065::Gmr/pUCP26-PA5065 | PAO1 with insertional mutation in PA5065, expressing PA5065 in trans from pUCP26 lac promoter | This study |
| PA5065::Gmr/pUCP20-ubiBEc | PAO1 with insertional mutation in PA5065, expressing E. coli ubiB in trans from pUCP20 lac promoter | This study |
| E. coli | ||
| JM109 | endA1 gyrA96 hsdR17(rK− mK+), mcrB+recA1 relA1 supE44 thi-1 Δ(lac-proAB) F′(traD36 proAB lacIZΔM15) | 71 |
| SM10 | thi-1 thr leu tonA lacY supE recA RP4-2-Tc::Mu Kmr | 54 |
| TOP10 | mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacX74 recA1 deoR araD139 Δ(ara-leu)7697 galU | Invitrogen |
| Plasmids | ||
| pCR2.1-TOPO | Topoisomerase-linked T/A cloning vector; Apr Kmr | Invitrogen |
| pUCP20 | pUC18-derived broad-host-range vector; Apr Pipr | H. Schweizer |
| pUCP26 | pUC18-derived broad-host-range vector; Tcr | 64 |
| pUCGM | Source of Gmr cassette; Apr Gmr | 51 |
| pEX18Ap | Gene replacement vector; oriT+sacB+ Apr | 27 |
| pUCP26-PA5065 | Construct used for in-trans expression of PA5065 in PA5065::Gmr insertional mutant | This study |
| pUCP20-ubiBEc | Construct used for in-trans expression of E. coli ubiB in PA5065::Gmr insertional mutant | This study |
| pEX18Ap-PA5065::Gmr | Construct used for generation of PA5065::Gmr insertional mutation | This study |
| pEX18Ap-0240::Gmr | Construct used for generation of PA0240::Gmr insertional mutation | This study |
| pEX18Ap-3710::Gmr | Construct used for generation of PA3710::Gmr insertional mutation | This study |
| pEX18Ap-3782::Gmr | Construct used for generation of PA3782::Gmr insertional mutation | This study |