TABLE 1.
Strains and plasmids used in this work
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| RN4220 | Restriction-deficient transformation recipient | 40 |
| RN6390 | Wild-type laboratory strain related to 8325-4, prophage-cured strain from human isolate NCTC8325 harboring an 11-bp deletion in rsbU that regulates sigB activity by activating RsbV, a factor that competitively binds to the anti-sigma factor RsbW | 40 |
| ALC488 | sarA mutant of RN6390 with sarA::ermC | 31 |
| ALC1713 | sarR mutant of RN6390 with sarR::ermC | 30 |
| ALC1927 | sarS mutant of RN6390 with sarS::ermC | 9 |
| ALC1905 | sarT mutant of RN6390 with sarT::ermC | 49 |
| ALC2272 | sarU mutant of RN6390 with sarU::ermC | 32 |
| ALC2319 | sarV mutant of RN6390 with saV::ermC | 33 |
| AM 1080 | sarX mutant of RN6390 with sarX::ermC | 34 |
| AM 1090 | sarZ mutant of RN6390 with sarZ::ermC | This study |
| ALC2530 | mgrA mutant of RN6390 with mgrA::ermC | 33 |
| AM 1102 | rot mutant of RN6390 with rot::ermC | This study |
| RN6911 | agr mutant of RN6390 with agr::tetM | 40 |
| AM1126 | sarZ agr double mutant of RN6390 with sarZ::kan and agr::tetM | This study |
| AM1175 | sarZ mgrA double mutant of RN6390 with sarZ::kan and mgrA::ermC | This study |
| AM1176 | sarZ sarX double mutant of RN6390 with sarZ::kan and sarX::ermC | This study |
| AM1194 | AM1090 complemented with 1.6-kb sarZ fragment into the geh locus on the chromosome | This study |
| AM1390 | AM1090 with pSK236 containing 1.6-kb sarZ gene at BamHI and XhoI sites | This study |
| SH1000 | Functional rsbU derivative of 8325-4 rsbU+ | 19 |
| AM1397 | sarZ mutant of SH1000 sarZ::ermC | This study |
| AM1398 | AM1397 complemented with the sarZ gene into the geh locus on the chromosome | This study |
| AM1410 | mgrA mutant of SH1000 mgrA::ermC | This study |
| AM1412 | AM1410 with pSK236 containing 1.5-kb mgrA fragment at EcoRI site | This study |
| Newman | A human clinical isolate | 37 |
| AM1419 | mgrA mutant of Newman mgrA::ermC | This study |
| AM1422 | AM1419 with pSK236 containing 1.5-kb mgrA gene at EcoRI site | This study |
| Plasmids | ||
| pUC18 | E. coli cloning vector | |
| pCR2.1 | E. coli cloning vector for direct cloning of PCR product | Invitrogen |
| pCL52.2 | Temperature-sensitive E. coli-S. aureus shuttle vector | 26 |
| pCL84 | Single-copy integration E. coli-S. aureus shuttle vector | 25 |
| pSK236 | E. coli-S. aureus shuttle vector | 16 |
| pAM927 | pUC19 containing a 2.3-kb BamHI- XhoI fragment containing sarZ region at BamHI-SalI sites | This study |
| pAM945 | pUC19 containing ∼3.3-kbDNA fragment that has a deletion of a 191-bp AflII/SwaI fragment that includes promoter and N-terminal (31-residue) regions of the sarZ gene and an insertion of the 1.2-kb ermC gene | This study |
| pAM950 | pCL52.2 containing ∼3.3-kbDNA fragment sarZ::ermC from pAM945 | This study |
| pAM1055 | Same as pAM945, but the ermC gene was replaced with 1.2-kb kanamycin gene (kan) | This study |
| pAM975 | pSK236 with a 1.6-kb DNA fragment containing the sarZ gene with 280-bp upstream sequence | This study |
| pAM980 | pCL84 with a 1.6-kb DNA fragment containing the sarZ gene with 280-bp upstream sequence | This study |
| pAM1110 | pSK236 with a 1.5-kb DNA fragment containing the mgrA gene region |