TABLE 1.

Strains and plasmids used in this study

Strain or plasmidGenotype, relevant phenotype, and/or selection markeraSource or reference
Strains
    E. coli TOP10F mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74 recA1 deoR araD139 Δ(ara leu)7697 galU galK rpsL (Strr) endIA1 nupGInvitrogen
    E. coli B178W3110 galE sup+12
    E. coli MLK217B178 lpxL::mini Tn10, Tetr12
    E. coli MKM10MLK217, pMKM.11, Apr, TetrThis study
    E. coli MKM11MLK217, pMKM.21, Apr, TetrThis study
    E. coli MLK1067B178 lpxM::Ωcam14
    E. coli MKM13MLK1067, pMKM.11, Apr, CmrThis study
    F. tularensis subsp. holarctica strain 1547-57Wild-type, human isolate36
    F. tularensis MKM87Strain 1547-57, lpxL::aphA-3, KmrThis study
Plasmids
    pBluescript II SK(−)Apr, cloning vectorStratagene
    pCR2.1TOPOApr, Kmr, cloning vectorInvitrogen
    pCRspec2pCR2.1TOPO, aad(9), Apr, Kmr, SprThis study
    pFNLTP10grosacBpFNLTP9 derivative, LVS groEL promoter, sacB, temperature-sensitive origin of Francisella replication, Kmr, AprGift from T. Zhart
    pMKM.1F. tularensis FTT0232c in pCR2.1, Apr, KmrThis study
    pMKM.11pBluescript II SK(−), EcoRI fragment from pMKM.1 containing FTT0232c, KmrThis study
    pMKM.16pCR2.1TOPO, FTT0232c and flanking sequence without consensus sequence regions, Apr, KmrThis study
    pMKM.17pMKM.16, aphA-3, Apr, KmrThis study
    pMKM.2pCR2.1TOPO, FTT0231c, Apr, KmrThis study
    pMKM.21pBluescript II SK(−), EcoRI fragment from pMKM.1 containing FTT0231c, KmrThis study
    pMKM.219c7.5-kb plasmid containing F. tularensis temperature-sensitive origin of replication, LVS groEL promoter upstream of sacB, E. coli origin of replication, aad(9), SprThis study
    pMKM.223pMKM.219c, FTT0232c::aphA-3, Spr, KmrThis study
    pSPECRpCRBlunt (Invitrogen), aad(9), Spr57
    pUC18KaphA-3, Kmr33
  • a Spr, Apr, and Kmr, resistance to spectinomycin, ampicillin, and kanamycin, respectively.