TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Reference or source | Comment |
|---|---|---|
| S. aureus strains | ||
| RN4220 | 30 | Restriction-deficient strain that accepts DNA from E. coli |
| Newman | 11 | S. aureus clinical isolate |
| Newman ΔhrtA | 45 | Isogenic Newman hrtA mutant |
| E. coli strains | ||
| DH5α | Strain for use in all cloning steps | |
| BL21(DE3) | Novagen | Recombinant protein expression strain |
| E. coli plasmids | ||
| pET15b | Novagen | E. coli expression construct with N-terminal His6 tag |
| pET15b.hrtA | This study | Expression construct for N-terminally His6-tagged full-length HrtA |
| pET15b.hrtA:K45A | This study | pET15b.hrtA derivative with K45A mutation |
| pET15b.hrtA:R76A | This study | pET15b.hrtA derivative with R76A mutation |
| pET15b.hrtA:G145A | This study | pET15b.hrtA derivative with G145A mutation |
| pET15b.hrtA:G145T | This study | pET15b.hrtA derivative with G145T mutation |
| pET15b.hrtA:E167Q | This study | pET15b.hrtA derivative with E167Q mutation |
| S. aureus plasmids | ||
| pOS1 | 36 | Empty E. coli/S. aureus shuttle vector |
| phrtA | 45 | ΔhrtA complementation construct containing hrtA under control of its native promoter |
| phrtA-myc | This study | ΔhrtA complementation construct containing c-myc-tagged hrtA under control of its native promoter |
| phrtA-myc:K45A | This study | phrtA-myc derivative with K45A mutation |
| phrtA-myc:R76A | This study | phrtA-myc derivative with R76A mutation |
| phrtA-myc:G145A | This study | phrtA-myc derivative with G145A mutation |
| phrtAmyc:G145T | This study | phrtA-myc derivative with G145T mutation |
| phrtAmyc:E167Q | This study | phrtA-myc derivative with E167Q mutation |