TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Source or reference | Comments |
|---|---|---|
| S. aureus | ||
| RN4220 | 25 | A mutant of 8325-4 that accepts foreign DNA |
| RN6390 | 25 | Laboratory strain that maintains its hemolytic pattern when propagated on sheep erythrocyte agar (parental strain) |
| RN6911 | 26 | An agr mutant of RN6390 with a Δagr::tetM mutation |
| PC1839 | 4 | 8325-4 with a sarA::kan mutation |
| ALC184 | 7 | sarA mutant of RN6390 with pRN6735 and pI524 |
| ALC865 | 7 | RN6911 (agr mutant) with pALC862 |
| ALC1016 | This study | RN6390 with pALC1014 |
| ALC1342 | This study | A sarA mutant in which the sarA gene (nt 586 to 1107) (1) has been replaced by an ermC gene |
| ALC1794 | 9 | RN6390 with pALC1639 |
| ALC1927 | This study | A sarS mutant of RN6390 with an ermC gene into the EcoRI site of the sarS gene |
| ALC2009 | This study | ALC1927 complemented with pALC2010 |
| ALC2033 | This study | RN6390 with Δagr::tetM and sarS::ermC mutations |
| ALC2034 | This study | ALC1927 (sarS mutant) with pALC1639 |
| ALC2057 | This study | RN6390 with a sarA::kan mutation |
| ALC2067 | This study | RN6390 with sarS::ermC and sarA::kan mutations |
| ALC2115 | This study | ALC1927 (sarS mutant) with pALC1014 |
| E. coli | ||
| XL1-Blue | 21 | A host strain for cloning |
| BL21 | 21 | A host strain for the pET14b expression vector |
| Plasmids | ||
| pCR2.1 | Invitrogen | E. coli cloning vector for direct cloning of PCR products |
| pUC18 | 21 | E. coli cloning vector |
| pCL52.2 | 17 | A temperature-sensitive E. coli-S. aureus shuttle vector |
| pET14b | Novagen | Expression vector for E. coli |
| pI524 | 16 | S. aureus plasmid containing a β-lactamase repressor |
| pLC4 | 31 | A shuttle plasmid containing a promoterless xylE reporter gene |
| pRN6735 | 16 | A derivative of pC194 containing the bla promoter and two-thirds of the blaZ gene followed by a 1.5-kb RNAIII fragment lacking its promoter |
| pSK236 | 12 | A shuttle vector containing pUC19 at the HindIII site of pC194 |
| pALC672 | This study | pCR2.1 with a 161-bp sarA P3 promoter fragment |
| pALC862 | 7 | pSK236 containing the entire sarA locus with the sarA ORF and the triple promoter system |
| pALC1014 | This study | pLC4 containing a 158-bp spa promoter fragment (nt 17 to 174) (20) |
| pALC1639 | 9 | pLC4 (transcriptional fusion vector) with a 491-bp spa promoter fragment (nt 1 to 174 plus 319 bp upstream) (20) |
| pALC1883 | This study | pUC18 containing a 1.8-kb sarS fragment (nt 2925 to 1082 in contig 6207) |
| pALC1889 | This study | Temperature-sensitive shuttle plasmid pCL52.2 containing the ermC gene at the EcoRI site (nt 2616 to 2621) of the 1.8-kb sarS fragment |
| pALC2010 | This study | Shuttle plasmid pSK236 containing a 1.2-kb sarS fragment (nt 3459 to 2189 of contig 6207) |
| pALC2040 | This study | pCR2.1 with a 1,562-bp spa structural gene (nt 219 to 1780) (20). |
| pALC2043 | This study | pET14b containing the 750-bp sarS gene at the XhoI/BamHI site |