TABLE 1.
Bacterial strains and plasmids
| Strain or plasmid | Description | Reference or source |
|---|---|---|
| E. faecalis strains | ||
| OG1RF (*)a | Rifampin- and fusidic acid-resistant derivative of human oral cavity isolate OG1 | 8 |
| OG1SSp (*) | Spectinomycin- and streptomycin-resistant derivative of human oral cavity isolate OG1 | 8 |
| OG1RF ΔEfaRFI | OG1RF EfaRFI (OG1RF_11622-11621) deletion mutant | This study |
| OG1RF ΔEfaRFI pAT28 | OG1RF ΔEfaRFI containing shuttle vector pAT28 | This study |
| OG1RF ΔEfaRFI pM.EfaRFI (*) | OG1RF ΔEfaRFI containing pM.EfaRFI | This study |
| OG1RF ΔEfaRFI::EfaRFIrbs | OG1RF ΔEfaRFI with chromosomal integration of OG1RF_11622-11621 between ORFs OG1RF_11778 and OG1RF_11779 | This study |
| OG1RF ΔEfaRFI::EfaRFIpro (*) | ΔEfaRFI::EfaRFIrbs with putative OG1RF_11622 promoter knocked in to OG1RF_11622-11621 integration | This study |
| V583 | Vancomycin-resistant clinical isolate | 9 |
| V583 pM.EfaRFI (*) | V583 containing pM.EfaRFI | This study |
| T11 (*) | Urinary tract infection isolate | 75 |
| CH188 | Liver abscess isolate | 76 |
| E. coli strains | ||
| EC1000 | Cloning host, providing repA in trans | 77 |
| TOP10 | Cloning host | Invitrogen |
| StbL4 | Cloning host | Life Technologies |
| BW25113 | Keio Collection host | 31 |
| JW1944 | Keio Collection dcm mutant | 31 |
| BL21(DE3) | Expression host strain | Novagen |
| Plasmids | ||
| pAT28 | Shuttle vector for E. faecalis; confers spectinomycin resistance | 35 |
| pLT06 | Markerless exchange plasmid; confers chloramphenicol resistance | 33 |
| pGEX-6p-1 | E. coli expression vector | GE Healthcare |
| pET28a | E. coli expression vector | Novagen |
| pCF10 | cCF10-inducible conjugative plasmid | 37 |
| pWH01 | pLT06 containing 2,306-bp XbaI/PstI-digested fragment of OG1RF_11621-2 | This study |
| pWH03 | pLT06 containing 1,998 XbaI/PstI-digested fragment of EF2238-NotI-EF2239 | This study |
| pWH02 | pLT06 containing 1,989-bp BamHI/PstI-digested fragment of EF2239 and OG1RF_11622 with its upstream intergenic region | This study |
| pWH21 | pGEX-6p-1 containing 1,275-bp BamHI/NotI-digested fragment of OG1RF_11823 | This study |
| pWH51 | pET28a containing 1,005-bp NdeI/BamHI-digested fragment of OG1RF_10790 | This study |
| pM.EfaRFI | pAT28 containing 1,543-bp BamHI/EcoRI-digested OG1RF_11622 with its promoter | This study |
| pCom02 | pWH03 containing 2,129-bp NotI-digested fragment of OG1RF_11621-2 with putative OG1RF_11622 ribosome binding site | This study |
a
Strain names followed by (*) are resistant to ApeKI and TseI digestion (Fig. 4). All E. faecalis strains were tested for ApeKI and TseI sensitivity.