TABLE 1

Strains and plasmids used in this study

Strain or plasmidCharacteristicsaSource or reference
Strains
    S. aureus
        USA300 HOUCommunity-associated MRSA isolate cured of pUSA300-HOU-MRSA49
        RN4220Restriction-deficient transformation recipient56
        NE1769USA300 JE2 ppiB::Bursa; ppiB NARSA transposon mutantNARSA
        NE914USA300 JE2 tig::Bursa; tig NARSA transposon mutantNARSA
        NE710USA300 JE2 nuc::Bursa; nuc NARSA transposon mutantNARSA
        NE163USA300 JE2 aur::Bursa; aur NARSA transposon mutantNARSA
        NE95USA300 JE2 agrB::Bursa; agrB NARSA transposon mutantNARSA
        NE678USA300 JE2 plc::Bursa; plc NARSA transposon mutantNARSA
        RKC0085USA300 HOU ΔprsAThis work
        RKC0113USA300 HOU ppiB::Bursa; ppiB mutantThis work
        RKC0114USA300 HOU tig::Bursa; tig mutantThis work
        RKC0165USA300 HOU agrB::Bursa; agrB mutantThis work
        RKC0181USA300 HOU nuc::Bursa; nuc mutantThis work
        RKC0182USA300 HOU aur::Bursa; aur mutantThis work
        RCK0178USA300 HOU plc::Bursa; plc mutantThis work
        RCK0063USA300 HOU hupA::pAZ106::hupA-HAThis work
        RKC0129RKC0085 containing plasmid pRKC0126; prsA+ complement strainThis work
        RKC0135RKC0113 containing plasmid pRKC0131; ppiB+ complement strainThis work
        RKC0072USA300 HOU(pMK4)This work
    E. coli
        DH5Cloning strainInvitrogen
        BL21(DE3)/pLysSProtein expression strainPromega
Plasmids
    pMK4Gram-positive shuttle vector (Cmr)52
    pJB38Temperature-sensitive allelic exchange plasmid (Cmr)50
    pAZ106Suicide plasmid (Eryr)
    pET24dC-terminal His6 tag expression vectorNovagen
    pMALc5XN-terminal MBP fusion expression vectorNEB
    pRKC0078pJB38 containing DNA flanking prsA geneThis work
    pRKC0126pMK4 prsA-His6This work
    pRKC0131pMK4 ppiB-HAThis work
    pRKC0061pAZ106 hupA-HAThis work
    pRKC0212pMALc5X ppiBThis work
    pRKC0276pET24d nucThis work
  • a Cmr, chloramphenicol resistance; Eryr, erythromycin resistance.