TABLE 1.
Bacterial strains and plasmids used in this study
Antibiotic resistance markers are abbreviated as follows: KanR, kanamycin resistance; GmR, gentamycin resistance; TetR, tetracycline resistance; CamR, chloramphenicol resistance.
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli TOP10TM | F− mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 araD139 Δ(ara-leu) 7697 galU galK rpsL (Strr) endA1 nupG λ- | Invitrogen |
| E. coli BL21-CodonPlus® (DE3)-RP | F− ompT hsdS (rB− mB−) dcm+ Tetr galλ (DE3) endA [argU proL Camr] | Stratagene |
| P. aeruginosa PAO1 | WT | Ref. 79 |
| PAO1 Δpsl | Deletion of the psl promoter | Ref. 9 |
| PAO1 ΔpslG | pslG deletion in PAO1 | Ref. 8 |
| PAO1 PBADpsl | psl-araC-PBAD promoter replacement; expression of psl operon upon induction with l-arabinose | Ref. 9 |
| PAO1 PBADpsl ΔpelF | In-frame deletion of pelF in PBADpsl | This study |
| PAO1 PBADpsl ΔpelF ΔpslG | In-frame deletion of pslG in PBADpsl ΔpelF | This study |
| PAO1 PBADpsl ΔpelF ΔpslD | In-frame deletion of pslD in PBADpsl ΔpelF | This study |
| PAO1 PBADpsl ΔpelF pslGE165Q | Chromosomal mutation of PslG E165Q in PBADpsl ΔpelF | This study |
| PAO1 PBADpsl ΔpelF pslGE276Q | Chromosomal mutation of PslG E276Q in PBADpsl ΔpelF | This study |
| PAO1 PBADpsl ΔpelF pslGE165Q/E276Q | Chromosomal mutation of PslG E165Q/E276Q in PBADpsl ΔpelF | This study |
| Plasmids | ||
| pEX18Gm | Suicide cloning vector, GmR | Ref. 33 |
| pEX18Gm::ΔpelF | pelF deletion construct, GmR | This study |
| pEX18Gm::ΔpslG | pslG deletion construct, GmR | This study |
| pPSV39 | P. aeruginosa in trans overexpression plasmid, GmR | Ref. 80 |
| pPSV39::pslG(1–442) | PslG (1–442) overexpression vector | This study |
| pET28a | E. coli expression vector with N-terminal His6 tag, KanR | Novagen |
| pET28a::pslG(31–442) | PslG (31–442) overexpression vector | This study |
| pET28a::pslG(40–360) | PslG (40–360) overexpression vector | This study |
| pET28a::pslG(31–442) E165Q | PslG (31–442) containing E165Q mutation overexpression vector | This study |
| pET28a::pslG(31–442) E276Q | PslG (31–442) containing E276Q mutation overexpression vector | This study |
| pET28a::pslG(31–442) E165Q/E276Q | PslG (31–442) containing E165Q/E276Q mutation overexpression vector | This study |