Table 1.

Strains and plasmids

Strain or plasmidResistance phenotypea or genotypeDescription (reference or source)
E. faecium
 C68Apr Emr GmrSmr Tcr Vmr Clinical VanB-type VRE isolate (this study)
 D366Vmr ClinicalE. faecium isolate (47)
 GE-1Fusr Rifr Recipient strain (10)
 CV133Apr TcrVmr Fusr Rifr Transconjugant resulting from mating between C68 and GE-1 (this study)
 CV142Apr Tcr VmrFusr Rifr Transconjugant resulting from mating between C68 and GE-1 (this study)
E. faecalis
 V583Vmr VanB VRE strain isolated in St. Louis, Mo. (13)
 CH19Bla+ EmrGmr Merr SmrTcr Clinical isolate (37)
 JH2-7Fusr Rifr Recipient strain (19)
 OG1XRFFusrRifr Recipient strain (18)
E. coli
 DH5αF φ80dlacZ(lacZYA-argF)U169 endA1 recA1 Transformation recipient strain (16)
 DH10BF mcrA Δ(mrr-hsdRMS-mcrBC) φ80dlacZΔM15 ΔlacX74 deoR recA1 ara Δ139Δ(ara leu)7697 galU galK λ-rpsL endA1 nupG Transformation recipient strain (Bethesda Research Laboratories)
Plasmids
 pAMβ1Emr Broad-host range conjugative plasmid (45)
 pUC18Apr Cloning vector (Bethesda Research Laboratories)
 pACYC184Cmr Tcr Cloning vector (39)
 pCRIIAprKmr Cloning vector for PCR products (Invitrogen)
 pCWR370Cmr Tcr 8.5-kbvanB-containing BglII/HindIII fragment of E. faecium C68 cloned into pACYC184
 pCWR374Apr 4-kbEcoRV/HindIII fragment of pCWR370 cloned into PUC18
 pCWR403Apr 8-kb HincIII fragment from the chromosome of E. faecium C68 containing the left end of Tn5382 cloned into pUC18
 pCWR404Apr 1.4-kb RsaI subfragment of pCWR403, containing end of Tn5382, used for sequencing
 pCWR406Apr 9-kb vanB-containingEcoRI fragment of E. faecium D366 containing right end of Tn5382 cloned into pUC18
 pCWR407Apr 1.2-kb RsaI fragment ofE. faecium D366 chromosome containing left end of Tn5382, cloned into pUC18
  • a Apr, ampicillin resistant; Cmr, chloramphenicol resistant; Emr, erythromycin resistant; Gmr, gentamicin resistant; Kmr, kanamycin resistant; Merr, mercuric chloride resistant; Smr, streptomycin resistant, Tcr, tetracycline resistant; Vmr, vancomycin resistant.