TABLE 1 .
Bacterial strains and plasmids
| Strain or plasmid | Genotype, phenotype, or relevant characteristic(s)c | Reference or source |
|---|---|---|
| Strain | ||
| P. aeruginosa | ||
| PAO1 | Serotype O5; A+ B+ | Laboratory stock |
| PAO1waaL | waaL::Gmr derived from strain PAO1; A− B− | 14 |
| PAO1wzx | wzx::Gmr derived from strain PAO1; A+ B− | 10 |
| PAO1wzy | wzy::Gmr derived from strain PAO1; A+ B− | 11 |
| E. coli DH10B | F− araD139 Δ(ara leu)7697 ΔlacX74 galU galK rpsL deoR ϕ80dlacZΔM15 endA1 nupG recA1 mcrA Δ(mrr hsdRMS mcrBC) | Laboratory stock |
| Plasmid | ||
| pBluescript II SK(+) | Escherichia expression vector (Plac); Apr | Fermentas |
| pHERD26T | Pseudomonas expression vector (PBAD araC); Tetr | 47 |
| pMA632 | Plasmid encoding phoA-lacZα dual-reporter construct; Apr | 15 |
| pPLE01a | pBluescript II SK+ with phoA-lacZα from pMA632; Apr | This study |
| pWaldo-GFPd wzx | Plasmid carrying gfp-his8; Kmr | 46 |
| pHERD26T-wzx-GFP | pHERD26T carrying wzx-gfp-his8; Tetr | This study |
| pPLE01-wzxb | Full-length wzx (encoding aa 1-411) fused to phoA-lacZα; Apr | This study |
| pWaldo-wzx-GFP wzy | pWaldo carrying wzx-gfp-his8; Kmr | This study |
| pHERD26T-wzy-GFP | pHERD26T carrying wzy-gfp-his8; Tetr | This study |
| pPLE01-wzyb | Full-length wzy (encoding aa 1-438) fused to phoA-lacZα; Apr | This study |
| pWaldo-wzy-GFP waaL | pWaldo carrying wzy-gfp-his8; Kmr | This study |
| pHERD26T-waaL-GFP | pHERD26T carrying waaL-gfp-his8; TetR | This study |
| pPLE01-waaLb | Full-length waaL (encoding aa 1-401) fused to phoA-lacZα; Apr | This study |
| pWaldo-waaL-GFP | pWaldo carrying waaL-gfp-his8; Kmr | This study |
a
All targeted truncations for wzx, wzy, and waaL were cloned upstream of phoA-lacZα in pPLE01 and are represented in Fig. 1, 2, and 3.
b
All random truncation libraries for wzx, wzy, and waaL were generated using the respective full-length fusion vectors and are represented in Fig. 1, 2, and 3.
c
Superscript “+” or “−” after A or B denotes the presence or absence of the particular O polysaccharide, respectively. Resistance (r) is shown for gentamicin (Gm), ampicillin (Ap), tetracycline (Tet), and kanamycin (Km). aa, amino acids.