TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Genotype or characteristic(s) | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| RN4220 | NCTC8325-4 r− m+ | Kreiswirth et al. (20) |
| TY34 | Clinical isolate (eta+agr type III mecA+) | |
| TY825 | Clinical isolate (etb+agr type IV) | |
| TF5367 | TY34 Δeta::cat | |
| FK101 | TY34 TCS2 MW0199::pFK5 | This work |
| FK102 | TY34 lytSRMW0236::pFK6 | This work |
| FK103 | TY34 graRSMW0621::pFK7 | This work |
| FK104 | TY34 saeRSMW0668::pFK8 | This work |
| FK105 | TY34 TCS6MW1208::pFK9 | This work |
| FK106 | TY34 arlRSMW1305::pFK10 | This work |
| FK107 | TY34 srrABMW1446::pFK11 | This work |
| FK108 | TY34 phoPRMW1637::pFK12 | This work |
| FK109 | TY34 yhcSRMW1790::pFK13 | This work |
| FK110 | TY34 vraSRMW1825::pFK14 | This work |
| FK111 | TY34 agrCAMW1962::pFK15 | This work |
| FK112 | TY34 kdpDEMW2002::pFK16 | This work |
| FK113 | TY34 hssRSMW2282::pFK17 | This work |
| FK114 | TY34 nreCBMW2314::pFK18 | This work |
| FK115 | TY34 TCS16MW2545::pFK19 | This work |
| FK128 | TY34 ΔsarA::cat | This work |
| FK129 | TY34 sarA::catsarS::pFK2 | This work |
| FK130 | TY34 sarS::pFK21 | This work |
| FK131 | TY34 sigB::pFK20 | This work |
| FK132 | TY34 sarA::catsaeRSMW0668::pFK8 | This work |
| FK133 | TY34 sarA::catagrCA MW1962::pFK15 | This work |
| FK134 | TY34 sarS::pFK21 saeRS MW0668::pFK8 | This work |
| FK135 | TY34 sarS::pFK21 agrCAMW1962::pFK15 | This work |
| FK136 | TY34 sigB::pFK20 complemented with pFK22 | This work |
| FK137 | TY34 ΔsarA::cat complemented with pFK23 | This work |
| FK138 | TY34 sarS::pFK21 complemented with pFK24 | This work |
| FK200 | TY825 etb mutant pETB cured | This work |
| FK204 | TY825 saeRSMW0668::pFK8 | This work |
| FK206 | TY825 arlRSMW1305::pFK10 | This work |
| FK211 | TY825 agrCAMW1962::pFK15 | This work |
| FK216 | TY825 sigB::pFK1 | This work |
| FK217 | TY825 sarS::pFK2 | This work |
| FK218 | TY825 ΔsarA::cat | This work |
| E. coli | ||
| DH5α | F− φ80dlacZΔM15 Δ(lacZYA-argF)U169deoRrecA1endA1hsdR17(rK− mK+) phoAsupE44 λ−thi-1gyrA96relA1 | TaKaRa |
| Plasmids | ||
| pUC19 | E. coli cloning vector | TaKaRa |
| pHY300PLK | Shuttle vector between E. coli and S. aureus | TaKaRa |
| pWH1520 | 7.9-kbp xylose-inducible vector; PxylA′xylR Ampr in E. coli, Tetr in S. aureus | Rygus and Hillen (32) |
| pCL52.1 | 8.0-kbp temp-sensitive shuttle vector; Spcr in E. coli, Tetr in S. aureus | Sau et al. (34) |
| pCL15 | 7.0-kbp IPTG-inducible vector; Pspac, Ampr in E. coli, Cmr in S. aureus | Luong and Lee (24) |
| pKFT | 5.7-kbp temp-sensitive shuttle vector; Ampr Tetr in E. coli, Tetr in S aureus | This work |
| pKFC | 5.1-kbp temp-sensitive shuttle vector; Ampr in E. coli, Cmr in S aureus | This work |
| pFK2 | pKFT containing 554-bp fragment of sarS | This work |
| pFK3 | pKFT containing sarA::cat for deletion of the sarA gene | This work |
| pFK4 | pKFT containing 547-bp fragment of MW0018a | This work |
| pFK5 | pKFT containing 658-bp fragment of MW0199a | This work |
| pFK6 | pKFT containing 775-bp fragment of MW0236a | This work |
| pFK7 | pKFT containing 640-bp fragment of MW0621a | This work |
| pFK8 | pKFT containing 545-bp fragment of MW0668a | This work |
| pFK9 | pKFT containing 486-bp fragment of MW1208a | This work |
| pFK10 | pKFT containing 517-bp fragment of MW1305a | This work |
| pFK11 | pKFT containing 552-bp fragment of MW1446a | This work |
| pFK12 | pKFT containing 533-bp fragment of MW1637a | This work |
| pFK13 | pKFT containing 456-bp fragment of MW1790a | This work |
| pFK14 | pKFT containing 510-bp fragment of MW1825a | This work |
| pFK15 | pKFT containing 543-bp fragment of MW1962a | This work |
| pFK16 | pKFT containing 595-bp fragment of MW2002a | This work |
| pFK17 | pKFT containing 534-bp fragment of MW2282a | This work |
| pFK18 | pKFT containing 472-bp fragment of MW2314a | This work |
| pFK19 | pKFT containing 529-bp fragment of MW2545a | This work |
| pFK20 | pKFC containing 503-bp fragment of sigB | This work |
| pFK21 | pKFC containing 554-bp fragment of sarS | This work |
| pFK22 | pWH1520 containing sigB gene | This work |
| pFK23 | pWH1520 containing sarA gene | This work |
| pFK24 | pWH1520 containing sarS gene | This work |
a
Locus numbers are based on S. aureus strain MW2 (http://www.bio.nite.go.jp). Gene names are based on S. aureus MW2 and according to the first reference where the TCS is described in the text.