TABLE 1 .
Strains and plasmids used in the present study
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| TF2758 | Wild-type clinical isolate, biofilm positive | This study |
| ATCC 49775 | Wild-type clinical isolate, biofilm negative | ATCC |
| FK300 | Derivative of NCTC8325-4 (rsbU repaired) | Laboratory stock |
| RN4220 | Restriction-negative strain, NCTC8325-4 derivative | 32 |
| TF2758 pC001 | TF2758 complemented with pC001 | This study |
| TF2758 pC002 | TF2758 complemented with pC002 | This study |
| robsm strain | FK300 with a stop mutation in rob | This study |
| Δrob strain | FK300 Δrob | This study |
| Δrob pC001 strain | FK300 Δrob complemented with pC001 | This study |
| Δrob pC003 strain | FK300 Δrob complemented with pC003 | This study |
| Δrob pKAT strain | FK300 Δrob complemented with pKAT | This study |
| Δrob ΔicaADB strain | FK300 Δrob ΔicaADB | This study |
| Δrob ΔUpstream strain | FK300 Δrob ΔSAOUHSC_2894 ΔSAOUHSC_2895 ΔSAOUHSC_2896 | This study |
| Δrob ΔDownstream strain | FK300 Δrob ΔSAOUHSC_2898 ΔSAOUHSC_2899 ΔSAOUHSC_2900 | This study |
| Δrob ΔSAOUHSC_2898 strain | FK300 Δrob ΔSAOUHSC_2898 | This study |
| Δrob ΔSAOUHSC_2899 strain | FK300 Δrob ΔSAOUHSC_2899 | This study |
| Δrob ΔSAOUHSC_2900 strain | FK300 Δrob ΔSAOUHSC_2900 | This study |
| E. coli | ||
| DH5α | Cloning strain | TaKaRa |
| BL21(DE3) | Host for recombinant protein production | Novagen |
| Plasmids | ||
| pGEM-T Easy | Cloning vector | Promega |
| pKAT | E. coli-S. aureus shuttle vector | 36 |
| pC001 | Vector for complementation experiments; containing rob from FK300 cloned in pKAT | This study |
| pC002 | Vector for complementation experiments; containing icaR from FK300 cloned in pKAT | This study |
| pC003 | Vector for complementation experiments; containing rob from TF2758 cloned in pKAT | This study |
| pC004 | Vector for complementation experiments; containing SAOUHSC_2898 from FK300 cloned in pKAT | This study |
| pKFT | Vector for allele replacement | 34 |
| pET-28a(+) | E. coli expression plasmid | Novagen |
| pET-22b(+) | E. coli expression plasmid | Novagen |
| pET28a-rob | His-Rob expression plasmid | This study |
| pET22b-icaR | His-IcaR expression plasmid | This study |