TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Descriptiona | Source or reference |
|---|---|---|
| E. coli DH5α | F− φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk− mk+) phoA supE44 λ−thi-1 gyrA96 relA1 | Invitrogen |
| P. aeruginosa strains | ||
| PA103 | Wild-type strain; serogroup O11 | 18 |
| Wzz1 mutant | PA103 with Gmr interruption in wzz1 | This study |
| Wzz2 mutant | PA103 with Tcr interruption in wzz2 | This study |
| Double mutant | PA103 with interruption in both wzz genes | This study |
| Plasmids | ||
| Topo2.1 | TA PCR cloning vector; Apr Kmr | Invitrogen |
| pEX18Ap | Gene replacement vector; Apr Cbr | 14 |
| pEX18-wzz1 | wzz1 interrupted with Gmr at PstI site, cloned into EcoRI site of pEX18Ap | This study |
| pEX100T | Gene replacement vector; Apr Cbr | 28 |
| pEX100-wzz2 | wzz2 interrupted with Tcr at SmaI site, cloned into SmaI site of pEX100T | This study |
| pHP45ΩTc | Source of Tcr cassette | 9 |
| pUCGM | Source of Gmr cassette | 27 |
| pRK2013 | Helper plasmid used in triparental matings; Kmr | 10 |
| pMMB66HE | Broad-host-range Ptac expression vector; Apr Cbr | 11 |
| pMMB-wzz1 | wzz1 coding region inserted into BamHI and SalI sites of pMMB66HE | This study |
| pMMB-wzz2 | wzz2 coding region inserted into HindIII and EcoRI sites of pMMB66HE | This study |
a
Ap, ampicillin; Km, kanamycin; Cb, carbinicillin; Gm, gentamicin; Tc, tetracycline; r, resistance.