TABLE 2.

Bacterial strains and plasmids used in this study

Strain or plasmidRelevant genotype (comments)Source or reference
E. coli strains
    CFT073Pyelonephritis isolate, fim+ pap+ hly+13
    CFT073OFF-intermediateΔ IRL, fim invertible element locked off, Kanr, sacB geneThis work
    CFT073OFFΔ IRL, fim invertible element locked offThis work
    CFT073ONΔ IRL, fim invertible element locked onThis work
    CFT073nalNalidixic acid-resistant derivative of wild-type CFT073 (equally virulent as wild-type strain in CBA mouse model of ascending UTI)26
    AAEC 189ΔfimB-H ΔrecA endA4
    Top10F mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara-leu)7697 galU galK rspL (Strr) endA1 nupGInvitrogen
Plasmids
    pNG400594-bp fragment from primers 439Nhe and ΔSK1M, ΔIRL, in PCR-BluntII-TopoThis work
    pNG401530-bp fragment from primers 439Bam and LOFF, ΔIRL, in PCR-BluntII-TopoThis work
    pNG402426-bp fragment from primers DRA1 and LON, ΔIRL, in PCR-BluntII-TopoThis work
    pNG403594-bp (439Nhe/ΔSK1M) fragment from pNG400 cloned into the BglII-PstI sites of pNG401, ΔIRLThis work
    pNG404594-bp (439Nhe/ΔSK1M) fragment from pNG400 cloned into the BglII-PstI sites of pNG402, ΔIRLThis work
    pNG4051,124-bp fragment from pNG403 cloned into the NheI-BamHI sites of pHK184, ΔIRL (locked off)This work
    pNG407kan-sacB fragment from pIB279 cloned into the BglII site of pNG405, ΔIRL (locked off)This work
    pNG409pNG407 with a ΔpUC origin of replication, temperature-sensitive pSC101 originThis work
    pNG410kan-sacB fragment from pIB279 cloned into the BglII site of pNG412, ΔIRL (locked off)This work
    pNG411pNG405 with a ΔpUC origin of replication, temperature-sensitive pSC101 originThis work
    pNG4121,020-bp fragment from pNG404 cloned into the NheI-BamHI sites of temperature sensitive vector pHK184(ΔpUC origin), ΔIRL (locked on)This work
    pHK184Camr, temperature-sensitive pSC101 origin of replication, pUC origin of replicationIan Blomfield, un- published data
    pIB279Ampr, Kanr, sacB-Neor cassette5
    PCR-BluntII-TopopMB1 origin of replication, kanamycin- and zeocin-resistant, ccdB lethal gene for selection of insertsInvitrogen