TABLE 1.

Strains and plasmids

Strain or plasmidRelevant featuresaReference or source
S. aureus strains
    RN4220S. aureus intermediate host strain (restriction negative) for cloning (derived from strain 8325-4)6
    M1vraSR mutant of RN4220 with ermC insertion in the EcoRI site of vraS, erythromycin resistantThis study
    IL-AMRSA clinical strain2
Plasmids
    pGEM-TEscherichia coli vector, ampicillin resistance (blaZ)Promega
    pAW8S. aureus-E. coli shuttle vector, Tetr, intergenic region from pUC118, pAMα1 gram-positive ori and ColE1 ori from E. coliAkihito Wada
    pCMVβVector used as a source for lacZ geneClontech
    pALC80Used as a source for ermC gene; pSPT181 containing a 2.4-kb sar fragment carrying ermC gene, stable at 32°CAmbrose Cheung
    pCL52.1E. coli-S. aureus shuttle plasmid; gram-positive Ts ori from pE194; Spxr; Tetr gene from pT181Chia Lee (10)
    pVRASR-1pGEM-T containing cloned vraSR PCR fragmentThis study
    pVRSASR-ermpVRASR-1 containing ermC in the EcoRI site of vraSThis study
    pVRASR-erm-pCL52.1PstI-digested pVRASR-erm ligated with PstI-digested pCL52.1This study
    pVP1pAW8 containing a translational fusion between the vraSR operon fragment, VP1, from strain IL-A and lacZ from pCMVβThis study
    pVP1′pAW8 containing a translational fusion between the vraSR operon fragment, VP1′, from strain IL-A and lacZ from pCMVβThis study
    pVP2pAW8 containing a lacZ translational fusion between the vraSR operon fragment, VP2, from MRSA strain IL-A and lacZ from pCMVβThis study
    pVP2′pAW8 containing translational fusion between vraSR operon fragment, VP2′, from MRSA strain IL-A and lacZ from pCMVβThis study
    pVP-262pAW8 containing a translational fusion between the VP-262 fragment from the vraSR operon from MRSA strain IL-A and lacZ from pCMVβThis study
    pVP-462pAW8 containing a translational fusion between the VP-462 fragment from the vraSR operon from MRSA strain IL-A and lacZ from pCMVβThis study
    pPBP2-bpAW8 containing a translational fusion between the pbp2 operon fragment, PBP2-b, from MRSA strain IL-A and lacZ from pCMVβThis study
    pPBP2-cpAW8 containing a translational fusion between the pbp2 operon fragment, PBP2-c, from MRSA strain IL-A and lacZ from pCMVβThis study
  • a The lacZ fragment in all lacZ fusions begins at the 33rd codon; ori, origin of replication; Ts, temperature sensitive.