TABLE 1.
Bacterial strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Relevant characteristic(s) or sequencea | Reference(s) or source |
|---|---|---|
| Strains | ||
| E. coli | ||
| K-12 JM109 | Subcloning host for pET22b derivatives | 34 |
| BL21(DE3) | λDE3 lysogen, T7 RNA polymerase gene; expression host | 30 |
| S. aureus | ||
| NCTC8325 | Biofilm-positive, laboratory strain, lysogenic for bacteriophages φ11 and φ12 | 16 |
| Cowan 1 | MSSA, catheter infections by binding of fibrinogen and fibronectin | 6, 18 |
| Newman | MSSA | 18, 32 |
| Wood 46 | Protein A-deficient, alpha-toxin-producing strain | 6 |
| S. simulans 22 | Penicillin resistant, lysostaphin susceptible | 19 |
| S. epidermidis O-47 | Biofilm-positive, clinical isolate causing CVC-associated infections | 13, 28 |
| Plasmids | ||
| pET22b | C-terminal six-His-tagged expression vector, T7 lac promoter, β-lactamase gene (Ampr); in this study, without pelB leader tag | 30 |
| pETerΔ11 | pET22b + ORF 53 of φ11 (aa 1 to 490, 56.5 kDa) (I/II) | This study |
| pETendo/ami11 | pET22b + truncated ORF 53 of φ11 (aa 1 to 371, 43.2 kDa) (I/IX) | This study |
| pETendo11 | pET22b + truncated ORF 53 of φ11 (aa 1 to 180, 22.1 kDa) (I/V) | This study |
| pETendoCBD11 | pET22b + truncated ORF 53 of φ11 (Δaa 181 to 370, 35.2 kDa) (I/V; VI/VII) | This study |
| pETami11 | pET22b + truncated ORF 53 of φ11 (aa 180 to 371, 22.8 kDa) (VIII/IX) | This study |
| pETamiCBD | pET22b + truncated ORF 53 of φ11 (aa 180 to 490, 36.1 kDa) (VIII/II) | This study |
| pETerΔ12 | pET22b + ORF 49 of φ12 (aa 1 to 484, 55.3 kDa) (III/IV) | This study |
| Primer no. and name | ||
| I; phi11F | 5′ TTTGGATCCAATGAGTATCATCATGGAGGTG 3′ (BamHI) | |
| II; phi11R | 5′ GTCAAGCTTACTGATTTCTCCCCATAAGTCA 3′ (HindIII) | |
| III; phi12F | 5′ GTGGGATCCAATGTTGATAACAAAAAACCAA 3′ (BamHI) | |
| IV; phi12R | 5′ TAAAAGCTTAATCGTGCTAAACTTACCAAAAC 3′ (HindIII) | |
| V; endo11R | 5′ TTGAAGCTTAGCTGTTTCTTTTTTAGGT 3′ (HindIII) | |
| VI; CBD11F | 5′ ATGAAGCTTAAAATACCGGTTGCCACTGT 3′ (HindIII) | |
| VII; CBD11R | 5′ GTCCTCGAGACTGATTTCTCCCCATAAGTCA 3′ (XhoI) | |
| VIII; ami11F | 5′ GAACATATGAAGCCACAACCTAAAGCAGTAG 3′ (NdeI) | |
| IX; ami11R | 5′ CGGAAGCTTACCATCCATGTACGCCCTAA 3′ (HindIII) |
a
Roman numerals in parentheses are primer pairs used for amplification of ORFs (forward/reverse). Restriction site sequences are underlined, and the sites are given in parentheses. MSSA, methicillin-susceptible S. aureus; CVC, central venous catheter.