TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or characteristics | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus | ||
| RN4220 | Heavily mutagenized derivative of NCTC 8325-4 | 63 |
| SH1000 | NCTC 8325-4 with repaired rsbU gene | 64 |
| JM27 | SH1000 ftsH::tetL | 17 |
| NCTC 8325-4 Δspx | NCTC 8325-4 Δspx | 60 |
| NCTC 8325-4 spx complement | NCTC 8325-4 spx mutant complemented with spx | 60 |
| ALC4976 | SH1000 ΔclpB | 22 |
| ALC4977 | SH1000 ΔclpC | 22 |
| ALC4978 | SH1000 ΔclpL | 22 |
| ALC4979 | SH1000 ΔclpQ | 22 |
| ALC4980 | SH1000 ΔclpX | 22 |
| ALC4981 | SH1000 ΔclpY | 22 |
| ALC5105 | SH1000 ΔclpP | 22 |
| ALC6490 | SH1000 clpC mutant complemented with clpC | 22 |
| ALC6491 | SH1000 clpP mutant complemented with clpP | 22 |
| ALC7010 | SH1000 ΔmcsB | This work |
| ALC7011 | SH1000 ΔtrfA | This work |
| ALC7137 | SH1000 ΔyjbH | This work |
| ALC7272 | SH1000 trfA mutant complemented with trfA | This work |
| E. coli XL1-Blue | General cloning strain | Agilent |
| Plasmids | ||
| pMAD | E. coli/S. aureus shuttle plasmid with the ori pE194ts; bgaB Ampr Ermr | 38 |
| pEPSA5 | Xylose-inducible shuttle vector; Ampr Cmr | 37 |
| pSK236 | S. aureus/E. coli shuttle vector with pUC19 cloned into the HindIII site of pC194 | 65 |
| pALC1484 | pSK236 with promoterless gfp-uvr, a cycle 3 gfp allele | 66 |
| pALC6188 | pEPSA5 with mazE with a 3′ myc tag and sarA ribosome binding site | 22 |
| pALC6486 | pEPSA5 with axe1 with a 5′ myc tag and sarA ribosome binding site | 22 |
| pALC6489 | pEPSA5 with axe2 with a 5′ myc tag and sarA ribosome binding site | 22 |
| pALC6954 | pSK236 with a phage repressor promoter and codon-optimized venus | This work |
| pALC6955 | pSK236 with a phage repressor promoter and codon-optimized venus-VAA | This work |
| pALC7016 | pEPSA5 with codon-optimized venus-VAA and sarA ribosome binding site | This work |
| pALC7113 | pEPSA5 with spx with a 5′ myc tag and sarA ribosome binding site | This work |
| pALC7151 | pEPSA5 with trfA with a 5′ myc tag and sarA ribosome binding site | This work |
| pALC7257 | pMAD::trfA (sa0857) complement | This work |
| pALC7422 | pSK236 with a phage repressor promoter and trfA with a 5′ myc tag | This work |
| pALC7867 | pMAD::ΔtrfA (sa0857) | This work |
| pALC7868 | pMAD::ΔmcsB (sa0482) | This work |
| pALC7869 | pMAD::ΔyjbH (sa0860) | This work |
| pALC7803 | pALC1484 with the spx P1 promoter in EcoRI and XbaI sites | This work |
| pALC7804 | pALC1484 with the spx P2 promoter in EcoRI and XbaI sites | This work |
| pALC7901 | pSK236 with a phage repressor promoter and codon-optimized venus-SsrA | This work |