TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or phenotype | Source or reference |
|---|---|---|
| Strains | ||
| E. coli | ||
| BL21(DE3) | F−ompT lon hsdSB(rB− mB−) DE3 | Novagen |
| DB11 | F−met thi gal nal rif hsdR Ems | 16 |
| JM110 | F′ traD36 lacIq Δ(lacZ)M15 proA+B+/rpsL (Strr) thr leu thi lacY galK galT ara fhuA dam dcm supE44 Δ(lac-proAB) | American Type Culture Collection |
| 21566 | K-12 thr-1 leuB6 Δ(gpt-proA)66 his G4 argE3 thi-1 rfbD1 lacY1 ara-14 galK2 xyl-5 mtl-1 mgl-51 rpsL31 kdgK51 supE44 rff::Tn10-66 | 17 |
| S. aureus | ||
| Newman | CP5 positive | NCTC 8178 |
| Newman P15-1 | ermB-inactivated cap5P gene | This study |
| RN4220 | Capsule negative, restriction negative | 20 |
| Plasmids | ||
| pERMB | 2.2-kb HpaI-SalI fragment from pRN8078 in pGEM-7Zf+ | This study |
| pET-24a+ | E. coli expression vector (Kmr) | Novagen |
| pGEM-7Zf+ | E. coli cloning vector (Apr) | Promega |
| pJCL84 | 6.2-kb HindIII fragment containing 3′ end of cap5 sequence in pGEM-7Zf+ | 9 |
| pKBK7 | 1.2-kb PCR amplicon carrying cap5P in pET-24a+ | This study |
| pKBK10 | 1.9-kb HindIII-EcoRI fragment carrying cap5P in pUC19 | 9 |
| pKBK13 | 1.3-kb BamHI-BclI fragment from pERMB inserted into cap5P in pKBK10 | This study |
| pKBK15 | 3.3-kb PCR amplicon from pKBK13 in pTS1 | This study |
| pKBK25 | 1.2-kb PCR amplicon carrying mnaA in pUC19 | This study |
| pRN8078 | Tn551 (ermB) in temperature-sensitive plasmid | 13 |
| pTS1 | Temperature-sensitive shuttle vector (Apr Cmr) | 19 |
| pUC19 | E. coli cloning vector (Apr) | New England Biolabs, Inc. |