TABLE 1.
Bacterial strains and plasmids used in this work
| Strain or plasmid | Relevant genotype and characteristics | Reference or source |
|---|---|---|
| E. coli | ||
| BL21(DE3) | F−hsdS gal ompT, lysogen of DE3 carrying T7 polymerase gene | 58 |
| TG1 | supE Δ(lac-proAB) thi hsdRΔ5 (F′::traΔ36 proA+B+ lacIqZΔM15) | 39 |
| P. aeruginosa | ||
| PAO1 | Prototroph, chl-2 | B. Holloway |
| PAO1ΔP | PAO1 derivative, chromosomal deletion of xcpP | This work |
| PAG2 | PAO1 derivative, chromosomal deletion of xcpQ | A. de Groot |
| Plasmids | ||
| pT7.5 | E. coli vector, Apr, φ10 promoter | 59 |
| pMMB190 | Broad-host-range vector, Apr, lacUV5 promoter, lacZα | 43 |
| pMMB67HE | Broad-host-range vector, Apr, tac promoter | 18 |
| pACYC184 | E. coli vector, Tcr, Cmr | New England Biolabs |
| pJBS633 | E. coli vector, Tcr, Kmr | 10 |
| pGEX-2T | E. coli vector, Apr, GST gene fusion | AMRAD |
| pRK2013 | Kmr, ColE1 with Tra+ Mob+ | 13 |
| pKNG101 | Smr, oriR6K, oriTRK2, mobRK2, sacB R+ (suicide vector) | 32 |
| pHP45ΩHg | Hgr, Ω interposon | 11 |
| pMB4 | xcpQ gene in pMMB67HE | A. de Groot |
| pSB10, pSB58 | xcpP gene cloned in pMMB67HE or pUC19, Apr | This work |
| pSB95 | 510-bp xcpP internal deletion in pKNG101, Tcr, Smr | This work |
| pMG1, pMG2 | xcpP with an ATG→TGA or TTG→ATC change in pMMB67HE | This work |
| pSB60, pSB62 (Tc′XcpPp) | tc′-xcpP gene fusion in pJBS633 or pMMB190, Kmr or Apr | This work |
| pSB75 (XcpPΔX) | 3′ deletion in xcpP at the XhoI site in pMMB190 | This work |
| pSB94 (XcpPΔP) | 3′ deletion in xcpP at the PvuI site in pMMB190 | This work |
| pSB74 (XcpPΔM) | 3′ deletion in xcpP at the MscI site in pMMB190 | This work |
| pSB86 (XcpPΔ19) | xcpP with an internal deletion of 85 bp in pMMB67HE | This work |
| pSB82b (XcpPΔ20) | xcpP with an internal deletion of 273 bp in pMMB67HE | This work |
| pSB18 | PCR product encoding lasB signal sequence in pUC19, Apr | This work |
| pSB20 (lasB-′xcpP) | PCR product encoding ′XcpPp in pSB18 | This work |
| pSB25, pSB24 | lasB-xcpP fusion of pSB20 in pT7.5 or pMMB190 | This work |
| pSB46 (lasB-′xcpZ) | PCR product encoding ′XcpZp in pSB18 | This work |
| pSB51 | lasB-xcpZ fusion of pSB46 in pMMB190 | This work |
| pSB54 | PCR product encoding XcpP periplasmic domain in pGEX-2T | This work |
| pSB65 | xcpY gene in pGEX-2T | This work |