TABLE 1.

Strains and plasmids used in this study

Strain or plasmidGenotype or descriptionSource or reference
Strains
    S. aureus strains
        RN4220 (EBII13)Parent strain16
        SA178RI (EBII16)RN4220 containing T7 polymerase11
        tar gene integrants
            EBII43SA178RI tarIJ::pSAKO-ΔtarIJ (Ermr Kanr)11
            EBII48SA178RI tarIJ′::pSAKO-ΔtarIJ′ (Ermr Kanr)This study
            EBII93SA178RI tarK::pSAKO-ΔtarK (Ermr Kanr)This study
            EBII94SA178RI tarL::pSAKO-ΔtarL (Ermr Kanr)This study
            EBII105EBII101 tarK::pSAKO-ΔtarK (Ermr Specr Kanr)This study
        Noncomplemented deletion mutants
            EBII100SA178RI tarK::Erm (Ermr Kans)This study
            EBII101SA178RI tarL::Erm (Ermr Kans)This study
            EBII49SA178RI tarIJ′::Erm (Ermr Kans)This study
        Complemented gene integrants
            EBII50EBII43/pG164-tarIJ11
            EBII113EBII43/pG164-tarJThis study
            EBII114EBII43/pG164-tarIJThis study
            EBII162EBII43/pG164-168_tagDFThis study
            EBII174EBII105/pLI50-tarKThis study
            EBII175EBII105/pLI50-tarLThis study
            EBII176EBII105/pG164-W23_tarKLThis study
            EBII177EBII105/pG164-tarKLThis study
        Complemented deletions
            EBII56SA178RI tarIJ::Erm/pG164-tarIJ11
            EBII118SA178RI tarIJ::Erm/pG164-tarJThis study
            EBII178EBII101 tarK::Spec (Ermr Specr Kans)/pLI50-tarKThis study
            EBII179EBII101 tarK::Spec (Ermr Specr Kans)/pLI50-tarLThis study
            EBII180EBII101 tarK::Spec (Ermr Specr Kans)/pG164-tarKLThis study
            EBII181EBII101 tarK::Spec (Ermr Specr Kans)/pG164-W23_tarKLThis study
    E. coli strains
        Novablue (EB1)endA1 hsdR17(rK12 mK12+) supE44 thi-1 recA1 gyrA96 relA1 lac [F′ proA+B+ lacIqZΔM15::Tn10] TcrNovagen
        MC1061 (EB30)MC1061F′ lacIq lacZ::Tn5 (neoR) pro+7
Plasmids
    pUS19Source for Specr cassette2
    pMUTIN4Source for Ermr cassette29
    pSAKOE. coli replicating vector containing sacB [BamP] W29; Kanr11
    pG164-tarDE. coli-S. aureus shuttle vector for T7-based expression (Ampr Cmr)11
    pLI50E. coli-S. aureus shuttle vector for native expression (Ampr Cmr)19
    pBluescript SKII (+)Plasmid for cloning (Ampr)Stratagene
    pSAKO-ΔtarKPlasmid for single integration into tarK flank (Ermr Kanr)This study
    pSAKO-ΔtarKPlasmid for single integration into tarK flank (Specr Kanr)This study
    pSAKO-ΔtarLPlasmid for single integration into tarL flank (Ermr Kanr)This study
    pG164-168_tagDFpG164 containing tagD and tagF from B. subtilis 168This study
    pG164-W23_tarKpG164 containing tarK from B. subtilis W23This study
    pG164-W23_tarLpG164 containing tarL from B. subtilis W23This study
    pG164-W23_tarKLpG164 containing tarK from B. subtilis W23This study
    pG164-tarKLpG164 containing SACOL0238/0242 from S. aureus RN4220This study
    PG164-tarIJpG164 containing tarIJ (SACOL0240/0241) from S. aureus RN422011
    PG164-tarIJpG164 containing tarIJ′ (SACOL0236/0237) from S. aureus RN4220This study
    PG164-tarJpG164 containing tarIJ′ (SACOL0237) from S. aureus RN4220This study
    pLI50-tarKpLI50 containing SACOL0238 from S. aureus RN4220This study
    pLI50-tarLpLI50 containing SACOL0242 from S. aureus RN4220This study