TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Genotype or description | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus strains | ||
| RN4220 (EBII13) | Parent strain | 16 |
| SA178RI (EBII16) | RN4220 containing T7 polymerase | 11 |
| tar gene integrants | ||
| EBII43 | SA178RI tarIJ::pSAKO-ΔtarIJ (Ermr Kanr) | 11 |
| EBII48 | SA178RI tarI′J′::pSAKO-ΔtarI′J′ (Ermr Kanr) | This study |
| EBII93 | SA178RI tarK::pSAKO-ΔtarK (Ermr Kanr) | This study |
| EBII94 | SA178RI tarL::pSAKO-ΔtarL (Ermr Kanr) | This study |
| EBII105 | EBII101 tarK::pSAKO-ΔtarK (Ermr Specr Kanr) | This study |
| Noncomplemented deletion mutants | ||
| EBII100 | SA178RI tarK::Erm (Ermr Kans) | This study |
| EBII101 | SA178RI tarL::Erm (Ermr Kans) | This study |
| EBII49 | SA178RI tarI′J′::Erm (Ermr Kans) | This study |
| Complemented gene integrants | ||
| EBII50 | EBII43/pG164-tarIJ | 11 |
| EBII113 | EBII43/pG164-tarJ′ | This study |
| EBII114 | EBII43/pG164-tarI′J′ | This study |
| EBII162 | EBII43/pG164-168_tagDF | This study |
| EBII174 | EBII105/pLI50-tarK | This study |
| EBII175 | EBII105/pLI50-tarL | This study |
| EBII176 | EBII105/pG164-W23_tarKL | This study |
| EBII177 | EBII105/pG164-tarKL | This study |
| Complemented deletions | ||
| EBII56 | SA178RI tarIJ::Erm/pG164-tarIJ | 11 |
| EBII118 | SA178RI tarIJ::Erm/pG164-tarJ′ | This study |
| EBII178 | EBII101 tarK::Spec (Ermr Specr Kans)/pLI50-tarK | This study |
| EBII179 | EBII101 tarK::Spec (Ermr Specr Kans)/pLI50-tarL | This study |
| EBII180 | EBII101 tarK::Spec (Ermr Specr Kans)/pG164-tarKL | This study |
| EBII181 | EBII101 tarK::Spec (Ermr Specr Kans)/pG164-W23_tarKL | This study |
| E. coli strains | ||
| Novablue (EB1) | endA1 hsdR17(rK12− mK12+) supE44 thi-1 recA1 gyrA96 relA1 lac [F′ proA+B+lacIqZΔM15::Tn10] Tcr | Novagen |
| MC1061 (EB30) | MC1061F′ lacIqlacZ::Tn5 (neoR) pro+ | 7 |
| Plasmids | ||
| pUS19 | Source for Specr cassette | 2 |
| pMUTIN4 | Source for Ermr cassette | 29 |
| pSAKO | E. coli replicating vector containing sacB [BamP] W29; Kanr | 11 |
| pG164-tarD | E. coli-S. aureus shuttle vector for T7-based expression (Ampr Cmr) | 11 |
| pLI50 | E. coli-S. aureus shuttle vector for native expression (Ampr Cmr) | 19 |
| pBluescript SKII (+) | Plasmid for cloning (Ampr) | Stratagene |
| pSAKO-ΔtarK | Plasmid for single integration into tarK flank (Ermr Kanr) | This study |
| pSAKO-ΔtarK | Plasmid for single integration into tarK flank (Specr Kanr) | This study |
| pSAKO-ΔtarL | Plasmid for single integration into tarL flank (Ermr Kanr) | This study |
| pG164-168_tagDF | pG164 containing tagD and tagF from B. subtilis 168 | This study |
| pG164-W23_tarK | pG164 containing tarK from B. subtilis W23 | This study |
| pG164-W23_tarL | pG164 containing tarL from B. subtilis W23 | This study |
| pG164-W23_tarKL | pG164 containing tarK from B. subtilis W23 | This study |
| pG164-tarKL | pG164 containing SACOL0238/0242 from S. aureus RN4220 | This study |
| PG164-tarIJ | pG164 containing tarIJ (SACOL0240/0241) from S. aureus RN4220 | 11 |
| PG164-tarI′J′ | pG164 containing tarI′J′ (SACOL0236/0237) from S. aureus RN4220 | This study |
| PG164-tarJ′ | pG164 containing tarI′J′ (SACOL0237) from S. aureus RN4220 | This study |
| pLI50-tarK | pLI50 containing SACOL0238 from S. aureus RN4220 | This study |
| pLI50-tarL | pLI50 containing SACOL0242 from S. aureus RN4220 | This study |