TABLE 1.

Strains and plasmids

Strain or plasmidRelevant detailsaReference or source
Strains
    S. aureus
        8325NCTC 8325; 11-bp deletion in rsbU33
        8325-48325 derivative cured of prophages; 11-bp deletion in rsbU25
        SH1000Functional rsbU derivative of 8325-4; rsbU+25
        RN4220Restriction-negative derivative of 8325-432
        DU5972sdrCD::Tcr; isogenic mutant of 8325-447
        DU5875spa::Tcr; isogenic mutant of 8325-452
        UAMS-240sarA::Tcr; isogenic mutant of S. aureus RN63904
        ICA1ΔicaADBC::Tcr; isogenic mutant of 8325-415
        DU5883fnbA::Tcr, fnbB::Emr; isogenic mutant of 8325-421
        SRTA1srtA::Tcr; isogenic mutant of SH1000This study
        CLFA1clfA::Tcr; isogenic mutant of RN4220This study
        FNBPB1fnbB::Tcr; isogenic mutant of RN4220This study
        DU5944clfA::Tn917 clfB::Tcr45
        SH1000 clfAB fnbABclfA::Tn917 clfB::Tcr from DU5944; fnbA::Tcr fnbB::Emr from DU588321, 45
        BH1CCMRSA clinical isolate; biofilm positive; SCCmec type II; MLST type 8; CC849
        BH3(04)MRSA clinical isolatel; biofilm positive; SCCmec type II; MLST type 8; CC849
        BH4(04)MRSA clinical isolate; biofilm positive; SCCmec type II; MLST type 8; CC849
        BH10(03)MRSA clinical isolate; biofilm positive; SCCmec type IV; MLST type 22; CC2249
        BH18(04)MRSA clinical isolate; biofilm positive in BHI glucose medium; SCCmec type IV; MLST type 22; CC2249
        BH30(04)MRSA clinical isolate; biofilm positive; SCCmec type II; MLST type 8; CC849
        BH40(04)MSSA clinical isolate; biofilm positive; MLST type 1099; CC549
        BH48(04)MSSA clinical isolate; biofilm positive; MLST type 8; CC849
        BH49(04)MSSA clinical isolate; biofilm positive; MLST type 6; CC649
        BH51(04)MSSA clinical isolate; biofilm positive; MLST type 5; CC549
        DAR217MRSA reference isolate; biofilm positive; SCCmec type I; MLST type 228; CC555
        DAR22MRSA reference isolate; biofilm positive; SCCmec type III; MLST type 5; CC555
        DAR35MRSA reference isolate; biofilm positive; SCCmec type II; MLST type 8; CC855
        DAR113MRSA reference isolate; biofilm positive; SCCmec type IV; MLST type 22; CC2255
        DAR141MRSA reference isolate; biofilm positive; SCCmec type II; MLST type 38; CC3055
        DAR199MRSA reference isolate; biofilm positive; SCCmec II; MLST type 36; CC3055
        DAR70MRSA reference isolate; biofilm positive; SCCmec type II; MLST type 45; CC4555
    E. coli TOPOrecA1 endA1 lac [F′ proAB lac1q Tn10 (Tetr)]Invitrogen
Plasmids
    pCR-Blunt II-TOPOPCR cloning vector; Kmr AprInvitrogen
    pBluescriptCloning vector; AprStratagene
    pT1814.45-kb S. aureus plasmid containing tetA(K)30
    pBlue-tetpBluescript containing the tetA gene from pT181 on a 2,352-bp HindIII fragmentThis study
    pBT2Temperature-sensitive E. coli-Staphylococcus shuttle vector; Apr (E. coli) Cmr (Staphylococcus)5
    pLI50E. coli-Staphylococcus shuttle vector35
    pMADShuttle vector for allele replacement; Apr (E. coli) Emr (Staphylococcus); contains bgaB gene encoding a thermostable β-galactosidase1
    pMAD-CmpMAD derivative; Apr (E. coli) Cmr (Staphylococcus)This study
    pBT2-tet2,352-bp HindIII fragment from pBlue-tet cloned into the HindIII site of pBT2This study
    pBT2-bga2,803-bp SmaI-SwaI fragment containing the bgaB gene from pMAD cloned into the SmaI site of pBT2This study
    pSAsrtA14,570-bp PCR product containing the srtA gene amplified from S. aureus 8325-4 using primers SAsrt1 and SAsrt2 cloned into pCR-Blunt II-TOPOThis study
    pSAsrtA2BamHI-PstI fragment from pSAsrtA1 cloned into the BamHI-PstI sites of pBT2This study
    pSAsrtA32,361-bp XbaI fragment containing the tetA gene from pBT2-tet cloned into XbaI site in the srtA gene of pSAsrtA2This study
    pSAsrtA41,080-bp PCR product containing the srtA gene amplified from S. aureus 8325-4 using primers SAsrtAcomp1 and SAsrtAcomp2 cloned into pCR-Blunt II-TOPOThis study
    pSAsrtA51,092-bp EcoRI fragment containing the srtA gene from pSAsrtA4 cloned into the EcoRI site of pLI50This study
    pSAclfA14,420-bp PCR product containing the clfA gene amplified from S. aureus 8325-4 using primers SAclfA1 and SAclfA2 cloned into pCR-Blunt II-TOPOThis study
    pSAclfA22,236-bp SwaI-SmaI fragment containing the tetA gene from pBlue-tet cloned into AleI site in the clfA gene of pSAclfA1This study
    pSAclfA36,656-bp BamHI-XbaI fragment containing the clfA::Tcr allele from pSAclfA2 cloned into the BamHl-Nhel sites of pBT2-bgaThis study
    pSAfnbB13,100-bp PCR product containing the fnbB gene amplified from S. aureus 8325-4 using primers SAfnbB1 and SAfnbB2 cloned into pCR-Blunt II-TOPOThis study
    pSAfnbB22,236-bp SwaI-SmaI fragment containing the tetA gene from pBlue-tet cloned into SwaI site in the fnbB gene of pSAfnbB1This study
    pSAfnbB35,400-bp BamHI-XbaI fragment containing the fnbB::Tcr allele from pSAfnbB2 cloned into the BamHl-Xbal site of pBT2This study
    pSAfnbB42,803-bp SmaI-SwaI fragment containing the bgaB gene from pMAD cloned into the SmaI site of pSAfnbB3This study
    pFnBPA4Shuttle plasmid derived from pGEM-7Zf (Ampr) and pSK265 (Cmr) carrying the full length fnbA gene21
    pFnBPB4Shuttle plasmid derived from pBluescript (Ampr) and pSK265 (Cmr) carrying the full length fnbB gene21
    pRM8pFnBPA4 expressing FnBPA minus the B, C, and D domains40
    pRM1pFnBPA4 expressing FnBPA minus the A domain40
  • a MLST, multilocus sequence typing.