TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Genotype and relevant markersa | Source or reference |
|---|---|---|
| Strains | ||
| E. coli MC1061 | 11 | |
| S. aureus | ||
| NCTC8325 | Type strain of S. aureus | 33 |
| BB255 | NCTC8325, penicillinase plasmid cured | 6 |
| RN4220 | NCTC8325, penicillinase plasmid cured, hly mutant, restriction minus | 24 |
| AW302 | RN4220(pAW11) | This study |
| AW303 | RN4220(pPBPA) | This study |
| AW304 | AW303 pbpA::pΔPBPA | This study |
| AW305 | AW302(pAW12) | This study |
| AW306 | AW303(pAW12) | This study |
| AW307 | AW304(pAW12) | This study |
| Plasmids | ||
| pAW119 | Cloning shuttle vector, pMB1 ori for replication in E. coli, pAMα1 ori for replication in gram-positive organisms, Tetr | This study |
| pAW9 | Suicide vector in S. aureus, p15A ori for replication in E. coli, Tetr | This study |
| pAW10 | Small (3.9-kb) shuttle vector, p15A ori for replication in E. coli, pAMα1 ori for replication in gram-positive organisms, Tetr | This study |
| pAW11 | Cloning shuttle vector, pMB1 ori for replication in E. coli, pAMα1 ori for replication in gram-positive organisms, Emr | This study |
| pAW12 | Cloning shuttle vector, the same origins as pAW11, Kmr | This study |
| pPBPA | pAW11 with a KpnI-KpnI 3.1-kb fragment containing orf2 and pbpA | This study |
| pΔPBPA | pAW9 with a HindIII-HindIII 936-bp fragment containing a middle part of pbpA | This study |
| pORF2 | pAW9 with a Sau3AI-HindIII 993-bp fragment containing intact orf2 | This study |
a
Tetr, tetracycline resistant; Emr, erythromycin resistant; Kmr, kanamycin resistant; ori, origin.