Three clinical isolates of Staphylococcus aureus resistant to gentamicin and other aminoglycosides have been examined for antibiotic modifying enzymes. The strains contain a number of these enzymes, most of them similar to those commonly found in aminoglycoside-resistant gram-negative strains. All three strains (and a transductant derived from one of them) contain two enzymes mediating gentamicin resistance, an aminoglycoside 6'-N-acetyltransferase and a novel enzyme, gentamicin phosphotransferase.