TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype and/or phenotypea | Source or reference |
|---|---|---|
| P. aeruginosa | ||
| PAO1 | Wild-type laboratory strain | Lab collection |
| PA14 | Human clinical isolate; Rifr | 45 |
| JY2M | pilC mutant of PA14; Rifr Cbr | This study |
| JY5M | pilA mutant of PA14; Rifr Cbr | This study |
| JY11M | ybtQ mutant of PA14; Rifr Cbr | This study |
| JY15M | uvrD mutant of PA14; Rifr Cbr | This study |
| E. coli | ||
| DH5α | F−endA1 hsdR17 supE44 thi-1 recA1 gyrA96 relA1 Δ(lacZYA-argF)U169 λ− φ80 dlacZΔM15 | Gibco BRL |
| SM10 | thi-I thi leu tonA lacY supE recA::RP4-2-Tc::Mu; Kmr | Lab collection |
| Plasmids | ||
| pUC19 | Cloning vector; Apr | Lab collection |
| pEX18 | Suicide vector for P. aeruginosa; oriT+sacB+, gene replacement vector with MCS from pUC18; Apr | 23 |
| pGEM-T Easy | PCR cloning vector; Apr | Promega |
| pJY1 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY2 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY3 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY4 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY5 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY6 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY8 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY10 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY11 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY12 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY13 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY15 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY16 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY17 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY19 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY20 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY22 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY23 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY24 | pEX18 derivative carrying RDA product; Apr | This study |
| pJY25 | pEX18 derivative carrying RDA product; Apr | This study |
| pJYP25 | pGEM-T Easy with a 3.0-kbp fragment of pilABC gene cluster; Apr | This study |
| pJY2A | pGEM-T Easy with a 1,160-bp IPCR product of pilC; Apr | This study |
| pJY5A | pGEM-T Easy with a 1,030-bp IPCR product of pilA; Apr | This study |
| pJY15A | pGEM-T Easy with a 1,150-bp IPCR product of uvrD; Apr | This study |
| pJY2-1 | pGEM-T Easy with a 600-bp internal fragment of pilC; Apr | This study |
| pJY5-1 | pGEM-T Easy with a 500-bp internal fragment of pilA; Apr | This study |
| pJY11-1 | pGEM-T Easy with a 1.3-kbp internal fragment of ybtQ; Apr | This study |
| pJY15-1 | pGEM-T Easy with a 1.1-kbp internal fragment of uvrD; Apr | This study |
| pJY2M | pEX18 derivative carrying a 600-bp internal fragment of pilC; Apr | This study |
| pJY5M | pEX18 derivative carrying a 500-bp internal fragment of pilA; Apr | This study |
| pJY11M | pEX18 derivative carrying a 1.3-kbp internal fragment of ybtQ; Apr | This study |
| pJY15M | pEX18 derivative carrying a 1.1-kb internal fragment of uvrD; Apr | This study |
| pJYYBT | 2.1-kb EcoRI fragment containing the intact ybtQ gene cloned into pUC19 | This study |
a
Apr, ampicillin resistance; Cbr, carbenicillin resistance; Kmr, kanamycin resistance; Rifr, rifampin resistance.