TABLE 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant genotype or phenotype | Source or reference |
|---|---|---|
| E. coli BL21(DE3) | F−ompT lon hsdSB (rB−mB−) DE3 | Novagen, Inc. |
| S. aureus | ||
| Reynolds | CP5 positive | 9 |
| JLO22 | 727-bp deletion in cap5O gene of Reynolds, CP5 negative | This study |
| RN4220 | Capsule negative, restriction negative | 27 |
| Plasmids | ||
| pET-24a+ | E. coli expression vector (Kmr) | Novagen, Inc. |
| pJCL24 | 9.1-kb EcoRI fragment from S. aureus Reynolds (cap5H to cap5P) in pLI50 | 21 |
| pKBK4 | 1.3-kb PCR amplicon carrying cap5O in pUC19 | 13 |
| pKBK5 | 9.1-kb EcoRI fragment (cap5H to cap5P) from pJCL24 in pUC19 | This study |
| pKBK9 | 4.1-kb AvaI-EcoRI fragment (cap5M to cap5P) from pKBK5 in pUC19 | This study |
| pKBK9-2 | 727-bp HpaI deletion (Δcap5O) of pKBK9 | This study |
| pKBK22 | 3.4-kb BamHI-EcoRI fragment (cap5M to cap5P; Δcap5O) from pKBK9-2 in pTS1 | This study |
| pKBK24 | 2.4-kb PCR amplicon carrying cap5O and cap5 flanking sequences in pLI50 | This study |
| pLI50 | Shuttle vector (Apr Cmr) | 16 |
| pNB1 | 1.3-kb XbaI-EcoRI fragment (cap5O) from pKBK4 in pET-28a+ | This study |
| pTS1 | Temperature-sensitive shuttle vector (Apr Cmr) | 26 |
| pUC19 | E. coli cloning vector (Apr) | New England Biolabs, Inc. |