TABLE 1.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant genotype or characteristic(s)Source or reference
Strains
    E. coli
        DH5αF endA1 hsdR17(rK mK) thi-1 λ recA1 gyrA96 relA1 rhoA supE44 deoR φ80dlacZΔM15 Δ(lacZYA-argF)U169Invitrogen
    D. nodosus
        VCS1703ASerogroup G, transformable virulent isolate34
        JIR3885VCS1703A fimO Ω aphA-3 (fimO1)Natural transformation with pJIR2618
        JIR3886VCS1703A fimO Ω aphA-3 (fimO2)Natural transformation with pJIR2618
        JIR3889VCS1703A fimP Ωerm(B) (fimP1)Natural transformation with pJIR2773
        JIR3890VCS1703A fimP Ωerm(B) (fimP2)Natural transformation with pJIR2773
        JIR3895VCS1703A fimN ΩaphA-3 (fimN1)Natural transformation with pJIR2798
        JIR3896VCS1703A fimN ΩaphA-3 (fimN2)Natural transformation with pJIR2798
        JIR3910VCS1703A pilE Ωerm(B) (pilE1)Natural transformation with pJIR3088
        JIR3911VCS1703A pilE Ωerm(B) (pilE2)Natural transformation with pJIR3088
        JIR3913VCS1703A pilC Δ(405-2373) ΩaphA-3 (pilC1)Natural transformation with pJIR3132
        JIR3914VCS1703A pilC Δ(405-2373) ΩaphA-3 (pilC2)Natural transformation with pJIR3132
Plasmids
    pBluescript II SK(+)Apr lacZ+ cloning vectorStratagene
    pWSK29Apr lacZ+ low-copy-no. cloning vector79
    pUC18KpUC18 containing aphA-3 inserted into SmaI site, nonpolar cassette48
    pGEM7Zf(−)Apr lacZ+ cloning vectorPromega
    pJIR1532pBluescript II SK(+) containing tet(M) located between D. nodosus rrnA promoter and terminatorRecombinant
    pJIR2412pBluescript II SK(+) PstI Ωerm(B)60
    pJIR2614pJIR2412 XbaI/BamHIΩ(935-bp PCR product containing 5′ end of pilQ) XhoI/KpnI Ω(927-bp PCR product containing 3′ end of pilQ)Recombinant
    pJIR2617pBluescript II SK(+) XbaI/EcoRI Ω(2.1-kb PCR product containing fimO+)Recombinant
    pJIR2618pJIR2617 HpaI ΩaphA-3Recombinant
    pJIR2691pWSK29 containing tet(M) located between D. nodosus rrnA promoter and terminatorRecombinant
    pJIR2714pJIR2412 EcoRI/HindIII Ω(940-bp PCR product containing internal pilQ)Recombinant
    pJIR2769pBluescript II SK(+) XbaI/BamHI Ω(1.9-kb PCR product containing fimP+)Recombinant
    pJIR2770pGEM7Zf(−) XbaI/BamHI Ω(1.9-kb fimP from pJIR2769 XbaI/BamHI)Recombinant
    pJIR2773pJIR2770 HindII Ωerm(B)Recombinant
    pJIR2797pGEM7Zf(−) XbaI/EcoRI Ω(2.6-kb PCR product containing fimN+)Recombinant
    pJIR2798pJIR2797 EcoRV ΩaphA-3Recombinant
    pJIR3087pGEM7Zf(−) BamHI/XbaI Ω(2-kb PCR product containing pilE+)Recombinant
    pJIR3088pJIR3087 T4-filled HindIII Ωerm(B)Recombinant
    pJIR3124pUC18K XbaI/BamHI Ω(1.4-kb PCR product containing 3′ fragment of pilC)Recombinant
    pJIR3132pJIR3124 T4-filled EcoRI Ω(1.3-kb PCR product containing 5′ fragment of pilC)Recombinant