TABLE 1

Bacterial strains and plasmids used in this study

Strain or plasmidStrainRelevant genotype or characteristicaSource or reference
Strains
    E. coli DH5αRestriction-deficient DNA cloning strainGibco/BRL
    S. aureus
        RN42208325-4, r m+, restriction-defective strain which accepts foreign DNA49
        COL (Tets) (cured for plasmid)AJ717HA-MRSA, bla ΔmecR1 mecI mecR250
        MW2CA-MRSA, bla+ ΔmecR1 mecI mecR253
        Mu3HA-MRSA, hVISA, bla mecR1 mecI mecR252
        CYL316Derivative of RN4220 containing the integrase for insertion of pLC84 into the lipase gene51
        Newman ΔprsAAJ3Newman prsA::kan12
        NewmanΔprsA-CbAJ4Newman prsA::kan geh::pCL84_prsA12
        MW2ΔprsAAJ724MW2 prsA::kan, Φ80α transductant of Newman ΔprsAThis study
        Mu3ΔprsAAJ457Mu3 ΔprsA, markerless deletion using pKOR1This study
        COLΔprsAAJ728COL prsA::kan, Φ80α transductant of Newman ΔprsAThis study
        COLΔprsA-CAJ760COL prsA::kan, geh::pCL84_prsA, Φ80α transductant of Newman ΔprsA-CThis study
        COL_prsA_Nterm+CtermAJ843COL prsA::kan geh::pCL84_prsA_Nterm+CtermThis study
        COL_prsA_PPIaseAJ900COL prsA::kan geh::pCL84_prsA_PPIaseThis study
        COL_prsA_Nterm+PPIaseAJ1016COL prsA::kan geh::pCL84_prsA_Nterm+PPIaseThis study
        COL_prsA_NterAJ1020COL prsA::kan geh::pCL84_prsA_NtermThis study
        COL_prsA_CtermAJ1018COL prsA::kan geh::pCL84_prsA_CtermThis study
Plasmids
    pKOR1E. coli-S. aureus thermosensitive shuttle vector, Ampr Camr, ATc counterselection24
    pAJ342pKOR1_prsA_Mu3
    pCL84tetK; S. aureus geh locus integrating plasmid51
  • a Camr, chloramphenicol resistance; Ampr, ampicillin resistance.

  • b prsA-C, prsA complemented.