Fifteen of 22 clinical isolates of Enterococcus faecalis with high-level aminoglycoside resistance isolated in Greece, which were tested for mating ability, co-transferred pheromone response genes together with aminoglycoside resistance determinants to a sensitive recipient strain. Nine of them belonged to the same pulsotype, while the remaining six isolates were genetically unrelated. The prgB gene, which encodes aggregation substance, was detected in all the clinical isolates and transconjugants by both PCR and DNA hybridization but prgA, which encodes the surface exclusion protein, was only detected in two isolates, whereas it is present in most pheromone response plasmids from other sources.