Table 1.

Strains and plasmids used

Strain or plasmid Reference or source Comments
S. aureus
 RN422032Mutant of 8325–4 that accepts foreign DNA
 RN6390 32 Laboratory strain that maintains its hemolytic pattern when propagated on sheep erythrocyte agar (parental strain)
 RN6911 34 agr mutant of RN6390 with Δagr::tetM mutation
 ALC4889 sar mutant with sarA::ermCmutation
 ALC1713 This study sarR mutant of RN6390 with ΔsarR::ermC mutation
 Cowan I17 Laboratory strain
 DB 10 Clinical blood isolate previously used in adhesion and endocarditis studies
 Newman29 Laboratory strain
S. epidermidis Strain from collection at Utrecht University Hospital
S. haemolyticus Strain from collection at Utrecht University Hospital
S. saprophyticus Strain from collection at Utrecht University Hospital
E. coli
 XL-1 Blue 26 Host strain for cloning
 DH5α 26 Host strain for cloning
Plasmids
 pCR2.1 Invitrogen E. coli cloning vector for direct cloning of PCR products
 pBluescript Stratagene E. coli cloning vector
 pUC18 26 E. coli cloning vector
 pACYC177New England Biolabs E. coli cloning vector
 pCL52.1 24 Temperature-sensitive E. coli-S. aureus shuttle vector
 pET11b NovagenExpression vector for E. coli
 pALC926 This study pUC18 containing a 49-bp fragment upstream of the P2 promoter of the sar locus
 pALC1357 This study pET11b containing the 345-bp sarR gene at theNdeI/BamHI site
 pALC1361 This studypACYC177 with a ∼4-kb ClaI fragment containing thesarR region of RN6390
 pALC1627 This studypBluescript with a 2.5-kb EcoRI/ClaI fragment containing the sarR gene subcloned from pALC1361
 pALC1687 This study pBluescript with a 290-bp deletion of the sarR gene in pALC1627
 pALC1696 This studypCL52.1 with a 290-bp sarR deletion replaced by theermC gene at the EcoRV/SalI site