Table 1. Strains and plasmids used in this study.
| Strain or plasmid | Relevant genotypea | Reference or source |
|---|---|---|
| S. aureus strains | ||
| RN4220 | 8325-4 r-, initial recipient for modification of plasmids which are introduced into S. aureus from E. coli | NARSAb |
| NCTC8325 | HA-MSSA, wild-type | NARSA |
| Δrsp | 8325 strain deletion of rsp | This study |
| Crsp | rsp chromosomal complementation of Δrsp | This study |
| Δagr | 8325 strain deletion of agr | This study |
| ΔagrΔrsp | 8325 agr rsp double mutant | This study |
| N315 | HA-MRSA, agr-deficient | NARSA |
| N315Δrsp | N315 strain deletion of rsp | This study |
| Newman | HA-MSSA, α- and β-toxin-deficient | [41,42] |
| NewmanΔrsp | Newman strain deletion of rsp | This study |
| E. coli strains | ||
| Trans1-T1 | Clone host strain, F- ϕ80 (lacZ) ΔM15ΔlacX74 hsdR (rK- mK+) ΔrecA1398 endA1 tonA | TransGen |
| BL21(DE3) | Express strain, F- ompT hsdSB (rB- mB-) gal dcm (DE3) | TransGen |
| Plasmids | ||
| pBTs | Shuttle vector, temp sensitive, ampr cmr | [43] |
| pBTsΔrsp | pBTs derivative, for rsp deletion, ampr cmr | This study |
| pBTs-up-rsp-down | pBTs derivative, for rsp chromosomal complementation, ampr cmr | This study |
| pBTsΔagr | pBTs derivative, for agr deletion, ampr cmr | This study |
| pLI50 | Shuttle vector, ampr cmr | [44] |
| pLIrsp | pLI50 derivative, harbouring ORF of rsp and its promoter, ampr cmr | This study |
| pRMC2 | Shuttle vector, anhydrspetracycline inducible, ampr cmr | [45] |
| pRMCrsp | pRMC2 derivative, harbouring ORF of rsp, ampr cmr | This study |
| pOS1-lacZ | Shuttle vector, with lacZ ORF lacking first 6 amino acids, ampr cmr | [46] |
| pOSrsp | POS1-lacZ derivative, harbouring 301-bp region of rsp promoter and 18 bp of rsp coding sequence from strain NCTC8325, ampr cmr | This study |
| pOShla | POS1-lacZ derivative, harbouring 235-bp region of hla promoter and 18 bp of hla coding sequence from strain NCTC8325, ampr cmr | [47] |
| pOSpsmα | POS1-lacZ derivative, harbouring 308-bp region of psmα promoter and 18 bp of psmα coding sequence from strain NCTC8325, ampr cmr | This study |
| pOSpsmβ | POS1-lacZ derivative, harbouring 226-bp region of psmβ promoter and 18 bp of psmβ coding sequence from strain NCTC8325, ampr cmr | This study |
| pRSF-Duet | Expression vector with a hexa-histidine tag, kanr | Novagen |
| pRSF-Duet-Rsp | pRSF-Duet derivative, with ORF of rsp, kanr | This study |
ar−, restriction system negative; kanr, kanamycin resistant; ampr, ampicillin resistant; cmr, chloramphenicol resistant.
bNARSA, Network on Antimicrobial Resistance in Staphylococcus aureus.