TABLE 1.
Strains and plasmids used in this study
| Strain or plasmid | Resistance phenotypea | Description (reference or source) |
|---|---|---|
| E. faecalis strains | ||
| CH19 | Bla+ Emr Gmr Merr Smr Tcr | Clinical isolate (18) |
| CH116 | Bla+ Emr Gmr Merr Smr Tcr | Clinical isolate (18) |
| JH2-7 | Fusr Rifr | Recipient strain (13) |
| OG1XRF | Fusr Rifr | Recipient strain (12) |
| UV202 | Fusr Rifr | Recombination-deficient recipient strain (28) |
| CX19 | Bla+ Emr Gmr Merr Smr Tcr Fusr Rifr | Transconjugant resulting from mating between CH19 and JH2-7 (18) |
| CV34 | Bla+ Emr Gmr Merr Smr Tcr Fusr Rifr | Tranconjugant resulting from mating between CH116 and JH2-7 (this study) |
| CV123 | Bla+ Emr Gmr Merr Smr Tcr Fusr Rifr | Tranconjugant resulting from mating between CH19 and OG1XRF (this study) |
| CV61 | Bla+ Emr Gmr Merr Smr Tcr Fusr Rifr | Transconjugant resulting from mating between CH19 and JH2-7 (this study) |
| Plasmids | ||
| pUC18 | Ampr | Cloning vector (Bethesda Research Laboratories) |
| pACYC184 | Cmr Tcr | Cloning vector (22) |
| pCRII | Ampr Kmr | Cloning vector for PCR products (Invitrogen) |
| pDL412 | Smr | aadE gene probe (15) |
| pCWR81 | Ampr | 398-bp internal AluI-ClaI fragment of IS1216 cloned into pUC18 (20) |
| pCWR281 | Ampr | 4.3-kb EcoRI-ClaI fragment of CX19, containing the region between the end of Tn5381 and the internal IS1216 within Tn5385, cloned into pUC18 (this study) |
| pCWR303 | Ampr | 650-bp HindIII-ClaI fragment of CV34, containing the left Tn5385-chromosome junction, cloned into pUC18 (this study) |
| pCWR308 | Ampr Smr | 2.6-kb ClaI fragment from CV34, containing the intact streptomycin resistance determinant, cloned into pUC18 (this study) |
| pCWR318 | Ampr | 1.7-kb ClaI fragment from CV123, containing the left Tn5385-chromosome junction, cloned into pUC18 (this study) |
| pCWR343 | Cmr Tcr | 7-kb ClaI fragment of JH2-7, containing the target for Tn5385 insertion, cloned into pACYC184 (this study) |
| pCWR347 | Cmr Tcr | 3.1-kb HindIII fragment of CV34, containing the right Tn5385-chromosome junction, cloned into pACYC184 (this study) |
| pCWR349 | Ampr | Insert from pCWR347 cloned into pUC18 (this study) |
| pCWR351 | Ampr | 1-kb HindIII subfragment of pCWR343, containing the target for Tn5385 insertion, cloned into pUC18 (this study) |
| pCWR389 | Ampr | 5-kb HindIII fragment of CV61, containing the junction of the ends of Tn5385 across a single copy of IS1216, cloned into pUC18 (this study) |
a
Bla+, β-lactamase positive.