Table 1.

Bacterial strains and plasmids

Strain or plasmidGenotype and relevant markersaSource or reference
Strains
 E. coliMC106111
 S. aureus
  NCTC8325Type strain ofS. aureus33
  BB255NCTC8325, penicillinase plasmid cured6
  RN4220NCTC8325, penicillinase plasmid cured, hlymutant, restriction minus24
  AW302RN4220(pAW11)This study
  AW303RN4220(pPBPA)This study
  AW304AW303pbpA::pΔPBPAThis study
  AW305AW302(pAW12)This study
  AW306AW303(pAW12)This study
  AW307AW304(pAW12)This study
Plasmids
 pAW119Cloning shuttle vector, pMB1 ori for replication inE. coli, pAMα1 ori for replication in gram-positive organisms, TetrThis study
 pAW9Suicide vector in S. aureus, p15A ori for replication in E. coli, TetrThis study
 pAW10Small (3.9-kb) shuttle vector, p15A ori for replication in E. coli, pAMα1 ori for replication in gram-positive organisms, TetrThis study
 pAW11Cloning shuttle vector, pMB1 ori for replication in E. coli, pAMα1 ori for replication in gram-positive organisms, EmrThis study
 pAW12Cloning shuttle vector, the same origins as pAW11, KmrThis study
 pPBPApAW11 with aKpnI-KpnI 3.1-kb fragment containing orf2 andpbpAThis study
 pΔPBPApAW9 with aHindIII-HindIII 936-bp fragment containing a middle part of pbpAThis study
 pORF2pAW9 with a Sau3AI-HindIII 993-bp fragment containing intact orf2This study
  • a Tetr, tetracycline resistant; Emr, erythromycin resistant; Kmr, kanamycin resistant; ori, origin.