TABLE 1.

Strains and plasmids used

Strain or plasmidReference or sourceComment
S. aureus strains
    RN4220 30 Mutant of 8325-4 that accepts foreign DNA
    RN6390 30 Laboratory strain that maintains its hemolytic pattern when propagated on sheep erythrocyte agar (parental strain)
    RN6911 6 agr mutant of RN6390 with an Δagr::tetM mutation
    JC1413 43 atl mutant of S. aureus
    ALC1342Laboratory strainA sarA mutant with deletion of ORF3 and the sarA ORF and replaced with an ermC gene, Δorf3sarA::ermC
    ALC488 29 sarA mutant of RN6390 with sarA::ermC
    ALC1629Laboratory strainALC1342 with sarB (nt 1 to 1349 plus 180 bp of upstream sequence) integrated into the geh locus on the chromosome
    ALC1713 30 sarR mutant of RN6390 with ΔsarR::ermC
    ALC1905 41 sarT mutant of RN6390 with ΔsarT::ermC
    ALC2272 31 sarU mutant of RN6390 with ΔsarU::ermC
    ALC1893 9 sarS mutant of RN6390 with ΔsarS::ermC
    ALC2530 17 mgrA mutant of RN6390 with ΔmgrA::ermC
    ALC2531 17 ALC2530 complemented with mgrA gene into the geh locus on the chromosome
    ALC2319This study sarV mutant of RN6390 with a deletion of ORF of the sarV gene and replaced with an ermC gene
    ALC2375This studyRN6390 with pALC2366
    ALC2376This studyALC2272 with pALC2366
E. coli strains
    XL-1 Blue 28 Host strain for cloning
    Topo InvαF′InvitrogenHost strain for the TA cloning vector
Primers
    pCL52.2 30 Temperature-sensitive E. coli-S. aureus shuttle vector
    pCR2.1Invitrogen E. coli cloning vector for direct cloning of PCR products
    pSK236 31 Shuttle vector containing pUC19 cloned into the HindIII site of pC194
    pUC19 28 E. coli cloning vector
    pALC1484 18 Modified pSK236 shuttle vector with a promoterless gfpuvr reporter gene preceded by an S. aureus ribosome binding site
    pALC2231This studypUC19 containing a 2.2-kb EcoRVIHindIII fragment containing sarV gene
    pALC2280This studypCL52.2 containing a 2.84-kb DNA fragment which has a deletion of a 565-bp BsaBl/SphI fragment that includes complete ORF of the sarV gene and an insertion of the 1.2-kb ermC gene at the BsaBI/SphI sites
    pALC2366This studypSK236 with a 1.4-kb DNA fragment containing the sarV gene with a 295-bp upstream sequence at the EcoRI site
    pALC2489This studypALC1484 with a 273-bp promoter fragment of sarV fused with the gfpuvr reporter gene at the EcoRI/XbaI sites