TABLE 1.

Strains and plasmids used in this study

StrainSubspecies/alternative strain designation(s)/genotype/phenotypeSource/reference
F. tularensis a
    FSC155 (LVS1) F. tularensis subsp. holarctica, live vaccine strain, LVS (in strain collection since 1993)ATCC 29684,
    FSC458 (LVS2)Past tularemia vaccine, code no. NDBR 101, lot no. 11; vial reconstituted and used, in strain collection since June 2004The National Drug Company
    FSC200 F. tularensis subsp. holarcticaHuman ulcer, 1998, Sweden
    FSC352 F. tularensis subsp. holarcticaHare, 1974, Sweden
    FSC237 F. tularensis subsp. tularensis, SCHU S4Human ulcer, 1941, Ohio
    FSC472FSC155/pAL10, FSC155 expressing pilA in trans, TcrThis study
    FSC681FSC458/pAL10, FSC458 expressing pilA in trans, TcrThis study
    FSC688FSC458/pilA+, FSC458 expressing pilA in cisThis study
    FSC741FSC458/FTT0086+, FSC458 expressing FTT0086 in cisThis study
    FSC693FSC458/FTT0918+, FSC458 expressing FTT0918 in cisThis study
    FSC694FSC458/pilA+/FTT0918+, FSC458 expressing pilA and FTT0918 in cisThis study
    FSC767FSC200 /FTT0918, FTT0918 insertion mutantThis study
E. coli
    Top10FmcrA Δ(mrr-hsdRMS-mcrBC), φ80lacZΔM15 ΔlacX74 recA1 deoR araD139ara-leu)7697 galU galK rpsL (Smr) endA1 nupGInvitrogen
    S17-1λpir recA thi pro hsdR M +, <RP4:2-Tc:Mu:Km:Tn7>Tpr Smr 32
    BL21 Star (DE3)FompT hsdSB (rB mB) gal dcm rne-131 (DE3)Invitrogen
Plasmids
    pCR2.1-TOPOTopo cloning vector, Ampr KmrInvitrogen
    pCR4.0-TOPOTopo cloning vector, Ampr KmrInvitrogen
    pKK214GFP groELS promoter, Ft ori, p15A ori, GFP,b Tcr 1
    pDM4Suicide plasmid. sacB, mobRP4, oriR6K, Cmr 21
    pDMK2Suicide plasmid. sacB, mobRP4, oriR6K, KmrK. Kadzhaev, unpublished
    pFTKan1EZ-Tn5, pMOD-2 F. tularensis codon-optimized KanR clone sequence, F. tularensis groELS promoterT. Kawula, unpublished
    pET-30a(+)Cloning vector carrying an N-terminal His-tag/thrombin/S-tag/enterokinase configuration and an optional C-terminal His tag sequence, KmrNovagen
    pAL10 pilA gene cloned into NdeI and EcoRI sites of pKK214GFP, Tcr 7
    pAL11 pilA gene and additionally 1 kb upstream and downstream of pilA cloned into XhoI and SacI sites of pDM4, Cmr 7
    pEMS36FTT0086 gene and additionally 1 kb upstream and downstream of FTT0086 cloned into the XhoI and SalI sites of pDM4, CmrThis study
    pKK223The cis complementation construct for FTT0918 cloned into SaII and NotI sites of pDMK2, KmrThis study
    pKK224PvuII fragment generated by PCR using primers groELS/KanR/E-C cloned into blunt-ended pDM4, CmrThis study
  • a All Francisella strains are from the Francisella Strain Collection at FOI, Umeå, and each strain has been given a strain collection number.

  • b GFP, green fluorescent protein.