TABLE 2.
Strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| Escherichia coli | ||
| Trans T1 | Cloning strain | TransGen |
| BL21(DE3) | Expression strain | TransGen |
| Staphylococcus aureus | ||
| NCTC8325 | WT | NARSA |
| RN4220 | 8325-4, r− | NARSA |
| WT-pLI50 | NCTC8325, WT with plasmid pLI50 | This study |
| ΔsavRS | NCTC8325, savRS-deleted strain | This study |
| ΔsavRS-pLI50 | savRS-deleted strain with plasmid pLI50 | This study |
| ΔsavRS-pLIsavRS | Complementary strain of ΔsavRS, ΔsavRS with pLIsavRS | This study |
| Plasmids | ||
| pBTs | Shuttle vector, temp sensitive, Ampr Cmr | 42 |
| pBTsavRS | pBTs derivative, for savRS deletion, Ampr Cmr | This study |
| pBTefb-his | pBTs derivative, for His tag insertion in efb | This study |
| pET22b(+) | Expression vector with a hexahistidine tag, Ampr | Novagen |
| pET28a(+) | Expression vector with a hexahistidine tag, Kanr | Novagen |
| pET22savS | Protein SavS expression vector, Ampr | This study |
| pET28savR | Protein SavR expression vector with a hexahistidine tag, Kanr | This study |
| pET28savS | Protein SavS expression vector with a hexahistidine tag, Kanr | This study |
| pLI50 | Shuttle cloning vector, Ampr Cmr | Addgene |
| pLIsavRS | pLI50 with savRS and its promoter, Ampr Cmr | This study |
| pOS1 | Shuttle vector, with lacZ ORF lacking first 6 amino acids, Ampr Cmr | 43 |
| pOSsavRS | POS1 derivative, harboring 267-bp region of savRS promoter and 18 bp of savR coding sequence from strain NCTC8325, Ampr Cmr | This study |
| pOShla | POS1 derivative, harboring 300-bp region of hla promoter and 18 bp of hla coding sequence from strain NCTC8325, Ampr Cmr | This study |
| pOSefb | POS1 derivative, harboring 299-bp region of efb promoter and 18 bp of efb coding sequence from strain NCTC8325, Ampr Cmr | This study |