Table 1.

Strains and plasmids used in this study

Strain(s) or plasmidaRelevant characteristicsbSource or reference
Strains
S. aureusRN4220MSSA strain restriction negativeR. Novick
S. aureusCOLHomogeneous MRSA (MIC, 1,600 μg/ml)RU collectionc
S. aureusRUSA4COL Tn551 mutant with insertion in mecAgene (MIC, 3 μg/ml)15
 RNpPBP2iII-1 and -5RN4220 with Pspac-pbp2 fusion in the chromosome, transformed with pMGPIIThis study
 COLpPBP2iII-1 and -5COL with Pspac-pbp2 fusion in the chromosome, transformed with pMGPIIThis study
 RUSA4pPBP2iC-2 and -3RUSA4 with Pspac-pbp2 fusion in the chromosomeThis study
 COLpPBP2iC-2 and -3COL with Pspac-pbp2 fusion in the chromosomeThis study
Plasmids
 pSP64 E. coli plasmid, Apr Promega
 pSP64EIntegrational vector forS. aureus, Apr Emr 18
 pSPT181cThermosensitive E. coli-S. aureus shuttle vector, Apr Tcr Cmr S. Wu
 pDH88 Bacillus subtilis plasmid with IPTG-inducible Pspac promoter and lacI gene9
 pGC2 E. coli-S. aureus shuttle vector, Apr Cmr 24
 pMGPI S. aureus integrational vector with IPTG-inducible Pspacpromoter and lacI gene, AprEmr This study
 pMGPII S. aureusreplicative plasmid containing lacI gene, AprCmr This study
 pPBP2ipMGPI withpbp2 ribosome-binding site and first 171 codons fused to Pspac promoter, Apr Emr This study
 pPBP2iCpSP64 with pbp2 ribosome-binding site and first 171 codons fused to Pspac promoter, Apr Cmr This study
  • a The PBP2 constructs are underlined to indicate that the plasmids are integrated into the host cell's chromosome.

  • b Apr, ampicillin resistant; Emr, erythromycin resistant; Cmr, chloramphenicol resistant.

  • c RU, The Rockefeller University, Laboratory of Microbiology.