TABLE 1.
Strains and plasmids used in the study
| Strain or plasmid | Characteristic(s) | Source or reference |
|---|---|---|
| CSF41498 | Biofilm-positive cerebrospinal fluid isolate (Beaumont Hospital, Dublin, Ireland) | 9 |
| 1457 | Biofilm-positive isolate from infected central venous catheter (Hamburg, Germany) | 36 |
| rbf mutant | CSF41498 derivative; rbf::ermB Emr | This study |
| 1457 sarR | 1457 derivative; ΔsarR::tet Tcr | This study |
| CSF41498 sarR | CSF41498 derivative; ΔsarR::tet transduced from 1457; Tcr | This study |
| rbf sarR mutant | CSF41498 derivative; ΔsarR::tet rbf::ermB Tcr Emr | This study |
| pCR-Blunt II-TOPO | PCR cloning vector; Kmr Apr | Invitrogen |
| pLI50 | Escherichia coli-Staphylococcus cloning vector; Apr (E. coli) Camr (Staphylococcus) | 37 |
| pMal-c2x | E. coli MBP protein expression vector; Camr | NEB |
| pET30 EK/LIC | His tag cloning vector | Novagen |
| pEPSA5 | E. coli-Staphylococcus cloning vector; multiple cloning site downstream of xylose-inducible promoter | 17 |
| pBT2 | Temperature-sensitive E. coli-Staphylococcus shuttle vector; Apr (E. coli) Camr (Staphylococcus) | 16 |
| pUC19 | E. coli cloning vector; Apr | NEB |
| pROJ6448 | pRN5101::pC221 AluI-C; temperature-sensitive staphylococcal origin of replication | 38 |
| pSERB1 | 2,445-bp rbf fragment in pCR-Blunt II-TOPO | This study |
| pSERB2 | rbf gene in pSERB1 interrupted with ermB | This study |
| pSERB3 | rbf::erm from pSERB2 in shuttle vector pBT2 | This study |
| pSERB5 | 2,800-bp fragment containing rbf and its upstream region in pCR-Blunt II-TOPO | This study |
| pSERB6 | rbf fragment from pSERB5 cloned into pLI50 | This study |
| pEPrbf | rbf and its upstream region in pEPSA5 | This study |
| pLIsarR | sarR and its upstream region in pLI50 | This study |
| pTSERM6 | Promoterless rbf in pCR-Blunt II-TOPO | This study |
| pSERM6 | rbf from pTSERM6 downstream of malE in pMAL-c2x | This study |
| psarR-his | sarR gene in pET30 EK/LIC | This study |