TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Relevant genotype or description | Reference(s) or source |
|---|---|---|
| Escherichia coli DH5α | F− ϕ80lacZΔM15 Δ(lacZYA-argF)U169 recA1 endA1 hsdR17(rK− mK+) phoA supE44 thi-1 gyrA96 relA1 tonA | Life Technologies |
| P. aeruginosa strains | ||
| PAO1 | Wild type | B. H. Holloway |
| PAO1 ΔgcbA | gcbA (PA4843) allelic gene replacement in PAO1 | 39 |
| PAO1 ΔdipA | PA5017::ISphoA/hah Tetr | 36 |
| PAO1 ΔbdlA | ΔbdlA in PAO1 Kmr | 12 |
| PAO1 ΔwspF | wspF allelic exchange in PAO1 | 41 |
| PAO1 ΔsiaD | PA0169::ISphoA/hah Tetr | 42 |
| PAO1 ΔwspR | wspR allelic exchange in PAO1 | 43 |
| PAO1::PbdlA-bdlA-V5/His6 | Chromosomal expression of bdlA-V5/His6 in PAO1 | 38 |
| PAO1 ΔgcbA::PbdlA-bdlA-V5/His6 | Chromosomal expression of bdlA-V5/His6 in PAO1 ΔgcbA | This study |
| PAO1::PgcbA-gcbA-V5/His6 | Chromosomal expression of gcbA-V5/His6 in PAO1 | This study |
| PAO1 ΔgcbA::PgcbA-gcbA-V5/His6 | Chromosomal expression of gcbA-V5/His6 in ΔgcbA | This study |
| Plasmids | ||
| pRK2013 | Helper plasmid for triparental mating; mob tra Kmr | 44 |
| pJN105 | Arabinose-inducible gene expression vector; pBRR-1 MCS araC-PBAD Gmr | 45 |
| pMJT-1 | araC-PBAD cassette of pJN105 cloned into pUCP18; Ampr (carbapenem resistant) | 46 |
| pJN-gcbA | C-terminally V5/His6-tagged gcbA cloned into pJN105 at XbaI | 39 |
| pCTX-gcbA-V5/6×His | C-terminally V5/His6-tagged gcbA gene and upstream 500 bp cloned into pMini-CTX | 39 |
| CTX-PbdlA-bdlA-V5/6×His | C-terminally V5/His6-tagged bdlA gene and upstream 500 bp cloned into pMini-CTX | 38 |
| pJN-gcbA(GGAAF) | C-terminally V5/His6-tagged gcbA with mutation to convert GGEEF motif to GGAAF cloned into pJN105 | This study |
| pJN-PASa | PASa-encoding segment of bdlA cloned into pJN105 | 38 |
| pMJT-bdlA-PASbTarH | C-terminally V5/His6-tagged PASaTar of bdlA cloned into pMJT-1 | 38 |
| pJN-bdlA-V5/6 × His | C-terminally V5/His6-tagged bdlA cloned into pJN105 | 37 |
| pJN-dipA-V5/6 × His | C-terminally V5/His6-tagged dipA cloned into pJN105 | 36 |
| pJN-PA2133 | PA2133 cloned into pJN105 | 43 |
| pJN-bdlA-G31A-His | C-terminally His6-tagged bdlA with G31A mutation cloned into pJN105 | 37 |
| pMJT-bdlA-HA | C-terminally HA-tagged bdlA cloned into pMJT-1 | This study |
| pJN-bfiS | C-terminally V5/His6-tagged bfiS cloned into pJN105 | 47, 48 |