TABLE 3.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics | Source or reference |
|---|---|---|
| E. coli strains | ||
| DH5α | Plasmid free, Lac− | New England Biolabs |
| BL21(DE3) | IPTG-inducible T7 RNA polymerase | Invitrogen |
| S. aureus strains | ||
| RN4220 | Restriction deficient, prophage cured | 36 |
| RN4220::ftsH | Transposon insertion of ΦΝΞ 4818 in ftsH was transduced into RN4220 | This study |
| USA300-0114 | Clinical isolate | NARSAa |
| USA300-P23 | USA300-0114 without plasmids 2 and 3 | 37 |
| NE132 | SAUSA300_2640 (= mbtS) mutant from Nebraska Transposon Mutant Library | BEI Resources |
| NE1755 | SAUSA300_2641 mutant from Nebraska Transposon Mutant Library | BEI Resources |
| NE 373 | SAUSA300_2642 mutant from Nebraska Transposon Mutant Library | BEI Resources |
| USA300::mbtS | Transposon insertion of NE132 in mbtS was transduced into USA300-P23 | This study |
| USA300 ΔftsH | ftsH deletion mutant of USA300-P23 | 20 |
| USA300 ΔftsH::mbtS | Transposon insertion in mbtS in NE132 was transduced into USA300ΔftsH | This study |
| USA300 ΔftsH::2641 | Transposon insertion in SAUSA300_2641 in NE1755 was transduced into USA300 ΔftsH | This study |
| USA300 ΔftsH::2642 | Transposon insertion in SAUSA300_2642 in NE373 was transduced into USA300 ΔftsH | This study |
| Plasmids | ||
| pET28a | Protein expression vector | Novagen |
| pET28a-ftsH-Strep | pET28a containing the ftsH gene with Strep tag sequence at the C terminus | This study |
| pOS1 | E. coli-S. aureus shuttle vector, Cmr | 38 |
| pOS1-PmbtS-lacZ | lacZ gene fused to the promoter sequence of mbtS | This study |
| pYJ335 | E. coli-S. aureus shuttle vector with anhydrotetracycline-inducible promoter | 39 |
| pCL-ftsH-His6 | ftsH-His6 cloned in pCL55 | 20 |
| pYJ-ftsH-His6 | pYJ335 containing the ftsH gene with His tag sequence at the C terminus | This study |
a
NARSA, Network on Antimicrobial Resistance in Staphylococcus aureus.