TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| Strains | ||
| S. aureus LAC* | Erythromycin-sensitive derivative of MRSA strain LAC; clonal complex 8 | 29 |
| LAC* spa | Derivative of LAC* deficient in protein A; spa::Kanr | This study |
| LAC* sbi | Derivative of LAC* deficient in Sbi; sbi::Emr | 13 |
| LAC* spa sbi | Derivative of LAC* deficient in protein A and Sbi; spa::Kanr sbi::Emr | This study |
| LAC* PD | Protease-deficient derivative of LAC*; Δaur ΔsspAB Δscp spl::Emr | 46 |
| LAC* sbi lytM | Derivative of LAC* sbi deficient in LytM; ΔlytM | This study |
| LAC* spa sbi lytM | Derivative of LAC* spa sbi deficient in LytM; ΔlytM | This study |
| LAC* sbi [SpAΩSdrESS] | Derivative of LAC* sbi where the SpA sorting signal has been replaced with the sorting signal of SdrE by allelic exchange | This study |
| LAC* sbi lytM [SpAΩSdrESS] | Derivative of LAC* sbi lytM where the SpA sorting signal has been replaced with the sorting signal of SdrE by allelic exchange | This study |
| Newman sbi | Derivative of S. aureus strain Newman, NCTC 8178, clonal complex 8, deficient in Sbi; sbi::Emr | 13 |
| Newman spa sbi | Derivative of Newman deficient in protein A and Sbi; spa::Kanr sbi::Emr | 13 |
| Newman sbi lytM | Derivative of Newman sbi deficient in LytM; ΔlytM | This study |
| Newman sbi [SpAΩSdrESS] | Derivative of Newman sbi where the SpA sorting signal has been replaced with sorting signal of SdrE by allelic exchange | This study |
| E. coli DC10B | dam+ Δdcm ΔhsdRMS endA1 recA1 | 24 |
| Plasmids | ||
| pRMC2 | Anhydrotetracycline-inducible expression vector; Ampr Cmr | 23 |
| pSpA | Plasmid pRMC2 containing the full-length spa gene | This study |
| pSpAΔSS | Plasmid pSpA lacking DNA encoding the SpA sorting signal (residues 474 to 508) | This study |
| pSpAΩSdrESS | Plasmid pSpA where DNA encoding the SpA sorting signal (residues 474 to 508) has been replaced with DNA encoding the SdrE sorting signal (residues 1117 to 1154) | This study |
| pRMC2-sbi ΔD1D2 | Plasmid pRMC2 containing DNA encoding the Sbi signal sequence (residues 1 to 40) and the D3D4 domains (residues 153 to 253) | 13 |
| pD3D4-SpASS | Plasmid pRMC2 containing an in-frame fusion between DNA encoding Sbi D3D4 domains (residues 153 to 253) and DNA encoding the SpA sorting signal (residues 474 to 508) | This study |
| pD3D4-SdrESS | Plasmid pRMC2 containing an in-frame fusion between DNA encoding Sbi D3D4 domains (residues 153 to 253) and DNA encoding the SdrE sorting signal (residues 1117 to 1154) | This study |
| pSpAΩSdrESS-DS | Plasmid pSpAΩSdrESS containing 538 bp of sequence downstream of the spa gene | This study |
| pIMAY | Temperature-sensitive vector for allelic exchange; Cmr | 24 |
| pIMAY::ΔlytM | Plasmid for creating a lytM deletion mutant; carries 509 bp of DNA from upstream and 512 bp of DNA from downstream of the lytM gene amplified from LAC | This study |
| pIMAY::SpAΩSdrESS | Plasmid for replacing DNA encoding the sorting signal of SpA with DNA encoding the sorting signal of SdrE on the chromosome of LAC* | This study |