TABLE 1.
Strains and plasmids used in this study
| Strain(s) or plasmida | Relevant characteristicsb | Source or reference |
|---|---|---|
| Strains | ||
| S. aureus RN4220 | MSSA strain restriction negative | R. Novick |
| S. aureus COL | Homogeneous MRSA (MIC, 1,600 μg/ml) | RU collectionc |
| S. aureus RUSA4 | COL Tn551 mutant with insertion in mecA gene (MIC, 3 μg/ml) | 15 |
| RNpPBP2iII-1 and -5 | RN4220 with Pspac-pbp2 fusion in the chromosome, transformed with pMGPII | This study |
| COLpPBP2iII-1 and -5 | COL with Pspac-pbp2 fusion in the chromosome, transformed with pMGPII | This study |
| RUSA4pPBP2iC-2 and -3 | RUSA4 with Pspac-pbp2 fusion in the chromosome | This study |
| COLpPBP2iC-2 and -3 | COL with Pspac-pbp2 fusion in the chromosome | This study |
| Plasmids | ||
| pSP64 | E. coli plasmid, Apr | Promega |
| pSP64E | Integrational vector for S. aureus, Apr Emr | 18 |
| pSPT181c | Thermosensitive E. coli-S. aureus shuttle vector, Apr Tcr Cmr | S. Wu |
| pDH88 | Bacillus subtilis plasmid with IPTG-inducible Pspac promoter and lacI gene | 9 |
| pGC2 | E. coli-S. aureus shuttle vector, Apr Cmr | 24 |
| pMGPI | S. aureus integrational vector with IPTG-inducible Pspac promoter and lacI gene, Apr Emr | This study |
| pMGPII | S. aureus replicative plasmid containing lacI gene, Apr Cmr | This study |
| pPBP2i | pMGPI with pbp2 ribosome-binding site and first 171 codons fused to Pspac promoter, Apr Emr | This study |
| pPBP2iC | pSP64 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.