TABLE 1.
Strains and plasmids
| Strain or plasmid | Relevant characteristic(s) | Source or reference |
|---|---|---|
| Strain | ||
| S. epidermidis CSF41498 | Biofilm positive, cerebrospinal fluid isolate | This study |
| S. epidermidis RP62A | Biofilm positive | ATCC 35984a |
| S. epidermidis ICAR1-4 | icaR::Emr | This study |
| S. aureus RN4220 | Restriction-negative, intermediate host for plasmid transfer from E. coli to S. epidermidis | 23 |
| E. coli XL-1 Blue | recA1 endA1 lac [F′ proAB lac1q Tn10(Tetr)] | Stratagene |
| Plasmid | ||
| pEC5 | pBluescript KS+ derivative. Source of ermB gene (Emr). Apr | 5 |
| pBT2 | Temperature-sensitive E. coli-Staphylococcus shuttle vector. Apr (E. coli) Cmr (Staphylococcus) | 5 |
| pSTBlue | PCR cloning vector. Kmr Apr | Novagen |
| pSTicaR | 786-bp PCR product containing the icaR gene amplified from CSF41498 using primers ICAR1 and IPR2 in pSTBlue | This study |
| pGEM3Z | Cloning vector. Apr | Promega |
| pSER1 | 810-bp BamHI-HindIII fragment from pSTicaR containing icaR in pGEM3Z | This study |
| pSER2 | 1,227-bp EcoRI-ClaI ermB fragment from pEC5 cloned into Bpu10I site of icaR gene in pSER1 | This study |
| pSER3 | 2,037-bp BamHI-HindIII fragment containing icaR::Emr in pBT2 | This study |
| pSER4 | 810-bp BamHI-HindIII fragment from pSER1 containing icaR cloned into pBT2 | This study |
a
ATCC, American Type Culture Collection.