Table 1.

Strains and plasmids used in this study

Strain or plasmidRelevant genotype or phenotypeaReference or source
Strains
E. coli
  DH10B Fφ80dlacZΔM15 recA1 Gibco
S. aureus
  RN4220NCTC8325-4, r m+ (restriction minus, modification plus), rsbU38
  BB255Essentially the same as NCTC8325, rsbU4
  Newman ATCC25904; clinical isolate, high level of clumping factor, rsbU+24
  MSSA1112Clinical isolate, bla rsbU+25
  GP266 RN4220, rsbU+sigB1(Am) Tcr6
  GP268 BB255,rsbU+V+W+sigB+ Tcr30
  IK181 BB255, ΔrsbUVWsigB::erm(B) Emr40
  IK184 Newman, ΔrsbUVWsigB::erm(B) Emr40
  MB32 Newman, asp23+asp23p::pECasp23p-luc+ Emr30
  MB33 BB255, rsbU asp23+asp23p::pECasp23p-luc+ Emr5
  MB39 MSSA1112, ΔrsbUVWsigB::erm(B) EmrThis study
  MB49 GP268, asp23+asp23p::pECasp23p-luc+ TcrEmr30
  MB69 Newman, ΔrsbUVWsigB::erm(B)asp23+asp23p::pECasp23p-luc+ TcrEmr30
  MB70 MSSA1112, ΔrsbUVWsigB::erm(B)asp23+asp23p::pECasp23p-luc+ TcrEmrThis study
  MB73 MSSA1112,asp23+asp23p::pECasp23p-luc+ EmrThis study
  MB90 BB255, ΔrsbUVWsigB::erm(B)asp23+asp23p::pECasp23p-luc+ TcrEmr30
  MB94 BB255,rsbU+hld+hldP::luc+ TcrEmrThis study
  MB95 BB255, rsbU hld+hldp::pEChldp-luc+ EmrThis study
  MB97 Newman, hld+hldp::pEChldp-luc+ EmrThis study
  MB98 BB255, rsbU sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ EmrThis study
  MB100 Newman,sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ EmrThis study
  MB101 Newman,sigB1(Am) sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ Tcr EmrThis study
  MB102 BB255,rsbU+sigB1(Am)sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ Tcr EmrThis study
  MB103Newman, sigB1(Am) hld+hldp::pEChldp-luc+ TcrEmrThis study
  MB104 MSSA1112,hld+hldp::pEChldp-luc+ EmrThis study
  MB105 MSSA1112, sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ EmrThis study
  MB112 MSSA1112,sigB1(Am) hld+hldp::pEChldp-luc+ TcrEmrThis study
  MB113 MSSA1112,sigB1(Am) sar+sar-P1-P2-P3::pECsar-P1-P2-P3-luc+ Tcr EmrThis study
Plasmids
 pSP-luc+ Apr, firefly luciferase casette vector Promega
 pEC1 AprEmr, 1.45-kb ClaI erm(B) fragment of Tn551 in pUC18 9
 pTX15 TcrPxyl, staphylococcal origin of replication50
 pIK64 Tcr, pTX15 derivative, containingsigB under the control of the xylose-inducible promoterPxyl40
 pSPsarP1-P2-P3Apr, 867-bp PCR fragment of sar promoter from strain RN4220 in pSP-luc+ This study
 pSPhldpApr, 1-kb PCR fragment ofhld promoter from strain RN4220 in pSP-luc+This study
 pECsarP1-P2-P3-luc+Emr, 2.5-kb KpnI-EcoRIsarP1-P2-P3-luc+ fragment of pSPsarP1-P2-P3 in pECI, S. aureus integration vector that inserts into the sar promoters (sarP1-P2-P3) This study
 pEChldP::luc+Emr, 2.6-kb KpnI-EcoRIhldp-luc+ fragment of pSPhldP in pEC1, S. aureus integration vector that inserts into the hldpromoter (hldp) This study
  • a Abbreviations are as follows: Apr, ampicillin resistant; Emr, erythromycin resistant; Tcr, tetracycline resistant.