TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| 8325-4 | Wild-type cured of known prophages | 41 |
| RN4220 | Accepts foreign DNA (r− m+) | 30 |
| COL | Methicillin resistant | 29 |
| PC6911 | agrΔ::tetM Tcr in 8325-4 background | 4 |
| PC1839 | sarA::km Kmr in 8325-4 background | 5 |
| LH9 | ebh::pAZ4353 Emr in 8325-4 background | This work |
| LH10 | ebh::pMUTHEX in 8325-4 background | This work |
| LH11 | ebh::pMUTHEX in PC6911 background | This work |
| LH12 | ebh::pMUTHEX in PC1839 background | This work |
| LH17 | ebh::pAZ4353 Emr in COL background | This work |
| SRC001 | ebh::pMUTHEX in COL background | This work |
| E. coli | ||
| DH5α | φ80dlacZΔM15 recA1 endA1 gyrA96 thi-1 hsdR17 (rK− mK+) supE44 relA1 deoR Δ(lacZYA-argF)U169 | Promega |
| BL21(DE3) | F−ompT gal [dcm] [lon] hsdSB (rB− mB−) | Novagen |
| Plasmids | ||
| pAZ106 | Promoterless, transcriptional lacZ fusion vector; Apr (E. coli), Emr (S. aureus) | 27 |
| pAZ4353 | pAZ106-based vector containing a ∼1.6-kb BamHI/EcoRI-cut fragment of ebh; Apr (E. coli), Emr (S. aureus) | This work |
| pMUTIN4 | Promoterless, transcriptional lacZ fusion vector; Apr (E. coli), Emr (S. aureus) | 52 |
| pMUTHEX | pMUTIN4-based vector containing a ∼1.2-kb BamHI/HindIII-cut fragment of ebh, creating an ebh::lacZ fusion; Apr (E. coli), Emr (S. aureus) | This work |
| pET24-d | His6 tag overexpression vector; Kmr | Novagen |
| pETH2 | pET24-d containing ∼1-kb internal ebh fragment encoding H2 polypeptide; Kmr | This work |