TABLE 1.

Strains and plasmids used

Strain or plasmidReference or sourceComments
S. aureus
    RN422031Mutant of 8325-4 that accepts foreign DNA
    RN639031Laboratory strain that maintains its hemolytic pattern when propagated on sheep erythrocyte agar (parental strain)
    RN691120 agr mutant of RN6390 with an Δagr::tetM mutation
    ALC1342Laboratory strainA sarA mutant with deletion of open reading frame 3 and the sarA open reading frame and replaced with an ermC gene
    ALC171326 sarR mutant of RN6390 with ΔsarR::ermC
    ALC190536 sarT mutant of RN6390 with sarT::ermC
    ALC207136 sarT mutant ALC1905 carrying pSK236 with wild-type sarT gene
    ALC192711 sarS mutant of RN6390 with sarS::ermC
    ALC2272This study sarU mutant of RN6390 with a deletion of amino acids 1 to 153 of the N terminus of the sarU gene product and its replacement with an ermC gene
    ALC2380This studyRN6390 with pALC2360
    ALC2381This studyALC2272 with pALC2360
    ALC2601This studyRN6390 with pALC2591
    ALC2602This studyRN6390 with pALC2599
    ALC2604This studyRN6911 with pALC2591
    ALC2605This studyRN6911 with pALC2599
    ALC2607This studyALC1905 with pALC2591
    ALC2608This studyALC1905 with pALC2599
    ALC2609This studyALC2272 with pALC2591
    ALC2610This studyALC2272 with pALC2599
    ALC2714This studyRN6390 with pALC2707
    ALC2716This studyALC1905 with pALC2707
    ALC2717This studyALC2272 with pALC2707
E. coli
    XL-1 Blue25Host strain for cloning
    Topo InvαF′InvitrogenHost strain for the TA cloning vector
    BL21(DE3)pLysSNovagenHost strain for the pET expression system
Plasmids
    pCL52.234Temperature-sensitive E. coli-S. aureus shuttle vector
    pCR2.1Invitrogen E. coli cloning vector for direct cloning of PCR products
    pET14bNovagenExpression vector in E. coli
    pSK23619Shuttle vector containing pUC19 cloned into the HindIII site of pC194
    pUC1925 E. coli cloning vector
    pALC148418Modified pSK236 shuttle vector with a promoterless gfpuvr reporter gene preceded by an S. aureus ribosome binding site
    pALC1740This studypALC1484 with the hla promoter
    pALC174236pALC1484 with the agr P2 promoter
    pALC174318pALC1484 with the agr P3 promoter
    pALC1904This studypET14b containing the 345-bp sarT gene at the XhoI/BamHI sites
    pALC2208This studypCR2.1 containing a 2.3-kb PCR sarU-sarT fragment with flanking upstream and downstream sequences
    pALC2229This studypUC19 containing a 2.3-kb EcoRV-KpnI fragment from pALC2208 at the HincII and KpnI sites
    pALC2240This studypCL52.2 containing a 2.9-kb DNA fragment, which has a deletion of a 547-bp ClaI-HincII fragment that includes residue 1 to 153 of the N terminus of the sarU gene product and an insertion of the 1.2-kb ermC gene at the EcoRI site
    pALC2360This studypSK236 with a 1.5-kb DNA fragment containing the sarU gene with a 392-bp upstream sequence at the EcoRI site
    pALC2591This studypALC1484 with a 310-bp promoter fragment of the sarU gene fused with the gfpuvr reporter gene at the EcoRI and XbaI sites
    pALC2599This studypALC1484 with a 147-bp promoter fragment of the sarU gene fused with the gfpuvr reporter gene at the EcoRI and XbaI sites
    pALC2707This studypALC1484 with a 310-bp promoter fragment of sarT fused with the gfpuvr reporter gene at the EcoRI site