Table 2 Strains and plasmids used in this study
From: YsxC, an essential protein in Staphylococcus aureus crucial for ribosome assembly/stability
Strain | Relevant genotype/markers | Source |
|---|---|---|
Escherichia coli | ||
EL250 | F- mcr Δ(mrr-hsdRMS-mcrBC) ϕ 80 lacZ ΔM15 Δlac×74 recA1 deoR araD139 Δ(ara-leu) 7697 galU galK rpsL (StrR) endA1 nupG [λcl857 araC-PBADflpe] | [57] |
GL1299 | EL250/pGL411 | This study |
TunerTM(DE3) pLacI | F- ompT hsdSB(rB- mB-) gal dcm lacY1 (DE3) pLacI (Cam) | Novagen |
Staphylococcus aureus | ||
LC101 | RN4220 ysxC::TAP-tag | This study |
LC102 | SH1000 spa | This study |
LC103 | SH1000 spa ysxC::TAP-tag | This study |
LC107 | RN4220 Pspac ~ysxC ysxC+ | This study |
LC108 | SH1000 Pspac ~ysxC ysxC+ | This study |
LC109 | SH1000 Pspac ~ysxC ysxC+/pGL485 | This study |
RN4220 | Restriction deficient transformation recipient | [58] |
SH1000 | Functional rsbU+ derivative of 8325-4 | [63] |
SJF590 | 8325-4 spa::tet | [62] |
Plasmid | Relevant genotype | Source |
pBS1479 | CBP/Protein A tag | [27] |
pDG1513 | Tetracycline resistance gene (tet) | [55] |
pELC1 | pGL411 derivative with TAP-kan cassette in frame with 3' end of SH1000 ysxC | This study |
pELC4 | pETBlue-1-based ysxC His6 tag translational fusion | This study |
pELC6 | Tet-T-Pspac cassette upstream ysxC gene in pGL411 | This study |
pETBlue-1 AccepTor | 3'-dA overhang cloning plasmid vector for protein overexpression; ColE1 ori | Novagen |
pGL400 | Tet-T-Pspac cassette | This study |
pGL411 | pOB derivative containing SH1000 ysxC and flanking regions | This study |
pGL433 | TAP-tag-kan cassette | This study |
pGL485 | pMJ8426-based lacI pE194ori cat | This study |
pMAL7 | Kanamycin resistance gene (kan) | [61] |
pMJ8426 | lacI pE194ori | [26] |
pOB | Erythromycin/lincomycin resistance gene (ery); ColE1 ori | [54] |