TABLE 1.
Strains and plasmids used in this studya
| Strain or plasmid | Genotype or characteristic | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| DH5a | F－, Φ80d lacZΔM15, Δ(lacZYA-argF)U169, deoR, recA1, endA1, hsdR17(rK− mK+), phoA, supE44, λ－, thi-1, gyrA96, relA1 | Toyobo |
| BL21(DE3) | F－, ompT, hsdSB(rB－ mB－), gal(λcI857, ind1, Sam7, nin5, lacUV5-T7gene1), dcm(DE3) | Promega |
| DC10B | E. coli K12, Δdcm mutant | 49, 50 |
| S. aureus | ||
| MR23 | Clinical isolate of MRSA (ST380) | 27 |
| MR23ΔisaA | isaA was deleted from MR23 | This study |
| MR23ΔsceD | sceD was deleted from MR23 | This study |
| MR23ΔisaAΔsceD | isaA and sceD were deleted from MR23 | This study |
| MR23ΔmecA | mecA was deleted from MR23 | This study |
| MR10 | Clinical isolate of MRSA (ST5) | 27 |
| MR10ΔisaA | isaA was deleted from MR10 | This study |
| MR10ΔsceD | sceD was deleted from MR10 | This study |
| MR10ΔisaAΔsceD | isaA and sceD were deleted from MR10 | This study |
| USA300 | Clinical isolate of MRSA (ST8) | ATCC BAA1717 |
| USA300ΔisaA | isaA was deleted from USA300 | This study |
| USA300ΔsceD | sceD was deleted from USA300 | This study |
| USA300ΔisaAΔsceD | isaA and sceD were deleted from USA300 | This study |
| Plasmids | ||
| pET29a | E. coli expression vector; KmR | Novagen |
| pETsceD | sceD cloned in pET29a | This study |
| pKOR1 | An E. coli/S. aureus shuttle vector plasmid for knockout of S. aureus genes by allelic exchange; ApR, CmR | 48 |
| pCN51 | An E. coli/S. aureus shuttle vector plasmid; ApR, EmR | 54 |
| pCN51Cm | An E. coli/S. aureus shuttle vector plasmid; ApR, CmR | This study |
| pCNisaA | isaA cloned in pCN51Cm | This study |
| pCNsceD | sceD cloned in pCN51Cm | This study |
| pCNmecA | mecA cloned in pCN51Cm | This study |
a
KmR, kanamycin resistance; ApR, ampicillin resistance; EmR, erythromycin resistance; CmR, chloramphenicol resistance; ST, sequence type.