TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmida | Properties | Reference(s) or source |
|---|---|---|
| S. aureus strains | ||
| RF122 | Wild-type strain from bovine mastitis (TSST+ SEC+) | 12 |
| RF122-1 | tst::Tcr mutant | This study |
| RF122-2 | tst::Tcrsec::Emr mutant | This study |
| RF120 | SaPIbov-negative strain of same clonal type as RF122 | 12 |
| KB103 | recA mutant | 3 |
| RN4220 | Restriction/modification− | 21 |
| Plasmids | ||
| pJRF | sec gene (150-bp internal deletion) with 1.4-kb TaqI Emr insert from pE194 cloned into pUC18 | This study |
| pJRF101 | 6.5-kb HindIII fragment in pBluescript KS(+) | This study |
| pJRF102 | 4-kb HindIII fragment from RF122 containing left-hand junction cloned into pUC19 | This study |
| pJRF103 | 1-kb PCR product from RF120 containing chromosomal insertion site of SaPIbov cloned into pCR2.1-TOPO | This study |
| pJRFsec::Emr | pJRF with pTS2 (Cmr) cloned into HindIII site flanking sec gene | This study |
| pRN6684 | tst::tmn (Tcr) Emr | 31 |
| pCR2.1-TOPO | Vector for cloning PCR products | Invitrogen |
| pE194 | 1.4-kb EmrTaqI fragment | 16 |
| pTS2 | Cmr, ts rep; derived from pTV1ts, MCSb from pBluescript | C. O'Connell (personal communication); 14, 33 |
| pBluescript KS(+) | Phagemid derived from pUC19; MCS, vector for blue/white colony selection | Stratagene |
a
Strains RF120 and RF122 were previously shown to be of the same clonal type by random amplified polymorphic DNA typing, multilocus enzyme electrophoretic typing, and ribotyping analysis (11).
b
MCS, multiple cloning site.