Table 1.

Strains and plasmids used in this study

Strain or plasmidSource or referenceComments
S. aureus
 RN422025A mutant of 8325-4 that accepts foreign DNA
 RN639025Laboratory strain that maintains its hemolytic pattern when propagated on sheep erythrocyte agar (parental strain)
 RN691126An agr mutant of RN6390 with a Δagr::tetM mutation
 PC183948325-4 with asarA::kan mutation
 ALC1847 sarA mutant of RN6390 with pRN6735 and pI524
 ALC8657RN6911 (agr mutant) with pALC862
 ALC1016This studyRN6390 with pALC1014
 ALC1342This studyAsarA mutant in which the sarA gene (nt 586 to 1107) (1) has been replaced by an ermC gene
 ALC17949RN6390 with pALC1639
 ALC1927This studyA sarSmutant of RN6390 with an ermC gene into the EcoRI site of the sarS gene
 ALC2009This studyALC1927 complemented with pALC2010
 ALC2033This studyRN6390 with Δagr::tetM andsarS::ermC mutations
 ALC2034This studyALC1927 (sarS mutant) with pALC1639
 ALC2057This studyRN6390 with asarA::kan mutation
 ALC2067This studyRN6390 withsarS::ermC andsarA::kan mutations
 ALC2115This studyALC1927 (sarS mutant) with pALC1014
E. coli
 XL1-Blue21A host strain for cloning
 BL2121A host strain for the pET14b expression vector
Plasmids
 pCR2.1Invitrogen E. coli cloning vector for direct cloning of PCR products
 pUC1821 E. coli cloning vector
 pCL52.217A temperature-sensitive E. coli-S. aureus shuttle vector
 pET14bNovagenExpression vector for E. coli
 pI52416 S. aureusplasmid containing a β-lactamase repressor
 pLC431A shuttle plasmid containing a promoterless xylE reporter gene
 pRN673516A derivative of pC194 containing thebla promoter and two-thirds of the blaZ gene followed by a 1.5-kb RNAIII fragment lacking its promoter
 pSK23612A shuttle vector containing pUC19 at theHindIII site of pC194
 pALC672This studypCR2.1 with a 161-bp sarA P3 promoter fragment
 pALC8627pSK236 containing the entire sarAlocus with the sarA ORF and the triple promoter system
 pALC1014This studypLC4 containing a 158-bp spapromoter fragment (nt 17 to 174) (20)
 pALC16399pLC4 (transcriptional fusion vector) with a 491-bp spa promoter fragment (nt 1 to 174 plus 319 bp upstream) (20)
 pALC1883This studypUC18 containing a 1.8-kb sarS fragment (nt 2925 to 1082 in contig 6207)
 pALC1889This studyTemperature-sensitive shuttle plasmid pCL52.2 containing the ermC gene at the EcoRI site (nt 2616 to 2621) of the 1.8-kb sarS fragment
 pALC2010This studyShuttle plasmid pSK236 containing a 1.2-kb sarS fragment (nt 3459 to 2189 of contig 6207)
 pALC2040This studypCR2.1 with a 1,562-bp spastructural gene (nt 219 to 1780) (20).
 pALC2043This studypET14b containing the 750-bp sarS gene at theXhoI/BamHI site