Table 1.
Bacterial strains and plasmids used in this study.
| Strains or plasmids | Relevant properties | References or sources |
|---|---|---|
| E. coli STRAINS | ||
| DH5α | ΔlacU169 hsdR17 recA1 endA1 gyrA96 thiL relA1 | Taylor et al., 1993 |
| S. aureus STRAINS | ||
| RN4220 | Restriction-deficient S. aureus strain | Kreiswirth et al., 1983 |
| JE2 | Wild-type; developed from a CA-MRSA, USA300-FPR3757 | Fey et al., 2012 |
| ΔrelP | relP deletion mutant of JE2 | Pando et al., 2017 |
| ΔrelQ | relQ deletion mutant of JE2 | This work |
| ΔrelPQ | relPQ deletion mutant of JE2 | This work |
| PLASMIDS | ||
| pLI50 | E. coli-S. aureus shuttle plasmid; AmpR (E. coli); CmR (S. aureus) | Lee et al., 1991 |
| pKOR1 | ATc-inducible suicide mutagenesis vector | Bae and Schneewind, 2006 |
| pALC2073 | E. coli-S. aureus shuttle plasmid with xyl/tetO promoter; | Bateman et al., 2001 |
| pKOR1-ΔrelQ | relQ deletion plasmid | This work |
| pMN12 | relP ORF with its native promoter cloned in KpnI/XbaI site of pLI50 | This work |
| pMN13 | relQ ORF with its native promoter cloned in KpnI/SalI site of pLI50 | This work |
| pMN14 | relP promoter fused with relQ ORF and cloned in KpnI/SalI site of pLI50 | This work |
| pMN15 | relQ promoter fused with relP ORF and cloned in KpnI/XbaI site of pLI50 | This work |
| pMN18 | E. coli lacZ engineered ORF cloned in XbaI/HindIII sites of pLI50 with Bacillus subtilis spoVG ribosomal binding site. | This work |
| pMN19 | relP promoter region cloned in KpnI/XbaI sites of pMN18 | This work |
| pMN20 | SAUSA300_0906 promoter region cloned in KpnI/XbaI sites of pMN18 | This work |
| pMN21 | relQ upstream region (including SAUSA300_0906 promoter and ORF) cloned in KpnI/XbaI sites of pMN18 | This work |
| pMN25 | relQ ORF cloned in EcoRI sites of pALC2073 | This work |
| pMN26 | mecA ORF cloned in EcoRI sites of pALC2073 | This work |
AmpR, ampicillin resistance; CmR, chloramphenicol resistance.