TABLE 1.
Bacterial strains and plasmids used in this study
| Plasmid or strain | Description | Reference/source |
|---|---|---|
| Plasmids | ||
| pMAD | E. coli-S. aureus shuttle vector containing a thermosensitive origin of replication, bgaB | 17 |
| pMADΔtrx | pMAD containing up- and downstream regions of sacol1794 | This study |
| pMADΔmmpL | pMAD containing up- and downstream regions of sacol2566 | This study |
| pCN40 | E. coli-S. aureus shuttle vector | 33 |
| pCNtrx | pCN40 harboring sacol1794 gene and its promoter | This study |
| pCNtrxP70Q | pCNtrx harboring mutated sacol1794 gene that resulted in P70Q substitution in its product | This study |
| pCNmmpL | pCN40 harboring sacol2566 gene and its promoter | This study |
| pCNmmpLT172I | pCNmmpL harboring mutated sacol2566 gene resulted in T172I substitution in its product | This study |
| S. aureus strains | ||
| COL | Hospital-acquired methicillin-resistant S. aureus parent strain | NARSA |
| COLR | COL expressing resistance to oxadiazole antibiotics | This study |
| COLI | COL expressing intermediate resistance to oxadiazole antibiotics | This study |
| COLΔtrx | COL lacking sacol1794 | This study |
| COLΔmmpL | COL lacking sacol2556 | This study |
| RN4220 | Restriction-deficient derivative of NCTC 8325-4 | 34 |
| RN9594 | RN4220 containing pCN40 | 33 |
| RN4220Δtrx | RN4220 lacking sacol1794 | This study |
| RN4220ΔtrxΩ | RN4220Δtrx strain containing pCN40 (vector-only control) | This study |
| RN4220ΔtrxΩtrx | RN4220Δtrx strain complemented by pCNtrx | This study |
| RN4220ΔtrxΩtrxP70Q | RN4220Δtrx strain complemented by pCNtrxP70Q | This study |
| RN4220ΔmmpL | RN4220 lacking sacol2556 | This study |
| RN4220ΔmmpLΩ | RN4220ΔmmpL strain containing pCN40 (vector only control) | This study |
| RN4220ΔmmpLΩmmpL | RN4220ΔmmpL strain complemented by pCNmmpL | This study |
| RN4220ΔmmpLΩmmpLT172I | RN4220ΔmmpL strain complemented by pCNmmpLT172I | This study |
| E. coli strain | ||
| DH5α | supE44 ΔlacU169(Φ80lacZΔM15) hsdR17 recA1 endA1 gyrA96 thi-1 relA1 | 35 |