Table 1.

Strains and plasmids used in this study

Strain or plasmidRelevant genotypeaReference or source
Strains
E. coli
  DH10BFφ80dlacZΔM15 recA1 Gibco
  BL21 (DE3)F ompT gal [dcm] [lon] hsdS B (rB mB ); with DE3Novagen
S. aureus
  RN4220NCTC8325-4 r m+(restriction minus; modification plus); rsbU 16
  MSSA1112Clinical isolate; bla rsbU + 7
  NewmanATCC25904; clinical isolate; high level of clumping factor;rsbU + 6
  GP266RN4220;rsbU + sigB1(Am) Tcr This study
  GP268BB255;rsbU + V + W + sigB + Tcr 10
  MB33BB255; rsbU asp23 + asp23p::pECasp23p-luc+ Emr 10
  MB49GP268;asp23 + asp23p::pECasp23p-luc+ EmrTcr 10
  MB51RN4220;crtM::pBTcrtM Tcr This study
  MB118MB33; rsbU rsbW1Emr This study
  MB119MB33; rsbU rsbW2(Am) Emr This study
  MB127MB33;rsbU rsbW3 Emr This study
  MB128MB33; rsbU rsbW4 Emr This study
  MB130MB33; rsbU rsbW5(Am) Emr This study
  MB132MB33; rsbU rsbW6 Emr This study
  MB137MB33;rsbU sigB2 Emr This study
  MB138MB33; rsbU rsbW7 Emr This study
  MB140MB33; rsbU chrX1Emr This study
  MB148MB33; rsbU rsbW8 Emr This study
  MB158MB137;rsbU + sigB1(Am) EmrTcr This study
  MB159MB137; rsbU crtM::pBTcrtM EmrTcr This study
  MB161MB140;rsbU + V + W + sigB + Tcr Emr This study
  MB162MB140; rsbU + sigB1(Am) Emr Tcr This study
  MB163MB140; rsbU crtM::pBTcrtM EmrTcr This study
  MB213MB137;rsbU + V + W + sigB + Emr Tcr This study
  MB215MB119; rsbU + sigB1(Am) Emr Tcr This study
Plasmids
 pBC SK(+)Cmr;E. coli cloning vectorStratagene
 pBTTcr; 1.6-kb PCR fragment of tet(L) from pHY300PLK into Alw261-digested pBC SK(+)10
 pEC1Apr Emr; 1.45-kb Clalerm(B) fragment of Tn551 in pUC184
 pET-24b(+)Kmr; E. coli expression vectorNovagen
 pSP-luc+Apr; firefly luciferase casette vectorPromega
 pPG11AprTcr; 6.6-kb PstI-EcoRIsigB fragment from strain BB255, including a 252-bpMluI-BstXI fragment of rsbU from strain COL replacing the corresponding region of the sigBoperon from strain BB255 and carrying a 1.6-kb PCR fragment oftet(L) from pHY300PLK integrated into a bluntedBsp1191 site downstream of sigB in pUC1910
 pETrsbW Apr; 486-bp PCR fragment ofrsbW from strain BB255 in pET24b(+)This study
 pSPasp23p Apr; 1.1-kb fragment of theasp23 promoter from strain COL into pSP-luc+10
 pECasp23p-luo+Emr; 2.7-kbKpnI-EcoRI asp23p-luc+ fragment of pSPasp23p in pEC1, S. aureus integration vector that inserts into the asp23 promoter (asp23p) 10
 pBTasp23p-luc+Tcr; 2.7-kbKpnI-EcoRI asp23p-luc+ fragment of pSPasp23p in pBT, S. aureus integration vector that inserts into the asp23 promoter(asp23p) 10
 pBTcrtM Tcr; 510-bp PCR fragment of crtM in pBT, S. aureusintegration vector that inserts into crtM This study
  • a Abbreviations are as follows: Am, amber mutation; Apr, ampicillin resistant; chrX1, unmapped mutation; Cmr, chloramphenicol resistant; Emr, erythromycin resistant; Kmr, kanamycin resistant; Tcr, tetracycline resistant.