TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Description | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| RN4220 | Restriction-deficient strain; host for plasmid vectors | 36 |
| SP4221 | RN4220 containing pVE1C integrated in ssp locus | This study |
| RN6390B | Wild type; high protease, low Fn binding | 38 |
| RN6911 | agrΔ::tetM Tcr; agr-null isogenic derivative of RN6390B | 38 |
| SP6391 | sspA is inactivated by insertion of ermAB; Emr Cms | This study |
| SP6912 | Mutant sspA allele from SP6391 transduced into RN6911; Emr Tcr | This study |
| E. coli DH5α | Host strain for construction of recombinant plasmids | 22 |
| Plasmids | ||
| pBKS+ | E. coli cloning vector; Apr | 52 |
| pUC18 | E. coli cloning vector; Apr | 56 |
| pPQ126 | S. aureus plasmid; source of E194ts temperature-sensitive replication origin | 31 |
| pUC-V8 | 1.3-kb sspA PCR amplicon, cloned at HindIII and XbaI sites of pUC18; Apr | This study |
| pBS-E1 | 1.4-kb erm PCR amplicon, cloned in pBKS at BamHI; Apr, Emr | This study |
| pUC-VE | pUC-V8 digested with BsmI at a unique site in ssp, then converted to blunt ends and ligated to the 1.4-kb erm fragment from pBS-E1; Apr Emr | This study |
| pUC-E194C | PstI-XbaI fragment of pPQ126, containing the E194ts replication origin and chloramphenicol acetyl transferase (cat), cloned in pUC18; Apr Cmr | This study |
| pVE1C | 4.0-kb E194Cat fragment excised from pUC-E194C with PvuII and cloned at SmaI site of pUC-VE; Apr Emr Cmr | This study |