TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5α | F−endA1 hsdR17(rK− mK−) thi-1 λ−recA1 gyrA96 relA1 rhoA supE44 deoR φ80dlacZΔM15 Δ(lacZYA-argF)U169 | Invitrogen |
| D. nodosus | ||
| VCS1703A | Serogroup G, transformable virulent isolate | 34 |
| JIR3885 | VCS1703A fimO Ω aphA-3 (fimO1) | Natural transformation with pJIR2618 |
| JIR3886 | VCS1703A fimO Ω aphA-3 (fimO2) | Natural transformation with pJIR2618 |
| JIR3889 | VCS1703A fimP Ωerm(B) (fimP1) | Natural transformation with pJIR2773 |
| JIR3890 | VCS1703A fimP Ωerm(B) (fimP2) | Natural transformation with pJIR2773 |
| JIR3895 | VCS1703A fimN ΩaphA-3 (fimN1) | Natural transformation with pJIR2798 |
| JIR3896 | VCS1703A fimN ΩaphA-3 (fimN2) | Natural transformation with pJIR2798 |
| JIR3910 | VCS1703A pilE Ωerm(B) (pilE1) | Natural transformation with pJIR3088 |
| JIR3911 | VCS1703A pilE Ωerm(B) (pilE2) | Natural transformation with pJIR3088 |
| JIR3913 | VCS1703A pilC Δ(405-2373) ΩaphA-3 (pilC1) | Natural transformation with pJIR3132 |
| JIR3914 | VCS1703A pilC Δ(405-2373) ΩaphA-3 (pilC2) | Natural transformation with pJIR3132 |
| Plasmids | ||
| pBluescript II SK(+) | AprlacZ+ cloning vector | Stratagene |
| pWSK29 | AprlacZ+ low-copy-no. cloning vector | 79 |
| pUC18K | pUC18 containing aphA-3 inserted into SmaI site, nonpolar cassette | 48 |
| pGEM7Zf(−) | AprlacZ+ cloning vector | Promega |
| pJIR1532 | pBluescript II SK(+) containing tet(M) located between D. nodosus rrnA promoter and terminator | Recombinant |
| pJIR2412 | pBluescript II SK(+) PstI Ωerm(B) | 60 |
| pJIR2614 | pJIR2412 XbaI/BamHIΩ(935-bp PCR product containing 5′ end of pilQ) XhoI/KpnI Ω(927-bp PCR product containing 3′ end of pilQ) | Recombinant |
| pJIR2617 | pBluescript II SK(+) XbaI/EcoRI Ω(2.1-kb PCR product containing fimO+) | Recombinant |
| pJIR2618 | pJIR2617 HpaI ΩaphA-3 | Recombinant |
| pJIR2691 | pWSK29 containing tet(M) located between D. nodosus rrnA promoter and terminator | Recombinant |
| pJIR2714 | pJIR2412 EcoRI/HindIII Ω(940-bp PCR product containing internal pilQ) | Recombinant |
| pJIR2769 | pBluescript II SK(+) XbaI/BamHI Ω(1.9-kb PCR product containing fimP+) | Recombinant |
| pJIR2770 | pGEM7Zf(−) XbaI/BamHI Ω(1.9-kb fimP from pJIR2769 XbaI/BamHI) | Recombinant |
| pJIR2773 | pJIR2770 HindII Ωerm(B) | Recombinant |
| pJIR2797 | pGEM7Zf(−) XbaI/EcoRI Ω(2.6-kb PCR product containing fimN+) | Recombinant |
| pJIR2798 | pJIR2797 EcoRV ΩaphA-3 | Recombinant |
| pJIR3087 | pGEM7Zf(−) BamHI/XbaI Ω(2-kb PCR product containing pilE+) | Recombinant |
| pJIR3088 | pJIR3087 T4-filled HindIII Ωerm(B) | Recombinant |
| pJIR3124 | pUC18K XbaI/BamHI Ω(1.4-kb PCR product containing 3′ fragment of pilC) | Recombinant |
| pJIR3132 | pJIR3124 T4-filled EcoRI Ω(1.3-kb PCR product containing 5′ fragment of pilC) | Recombinant |