The genes encoding for beta-lactamase (bla+) and resistance to metallic ions (cadmium, mercury, lead, arsenate, and arsenite) were located in a 56.2-kilobase plasmid, pZA10, isolated from a clinical strain, Staphylococcus aureus 6344. This strain produced enterotoxin B and enterotoxin C1. Elimination of pZA10 by either sodium dodecyl sulfate or heat treatment (43 degrees C) resulted in the loss of the capability of the bacteria to produce both enterotoxin B and enterotoxin C1. A physical map of pZA10 was constructed with BamHI, SalI and BglII restriction endonucleases. Penicillin-resistant, enterotoxin B- and C1-producing cotransformants were isolated by transformation with pZA10 DNA with either S. aureus RN450 or cured S. aureus 6344 as recipients. The transferred plasmids exhibited genetic instability shown by changes in restriction pattern and molecular size, loss of plasmid DNA, and addition of chromosomal DNA. Enterotoxin B production was related to a 18.1-kilobase pZA10 fragment carried by such a rearranged plasmid. Chromosomal cointegration of bla+ with genetic determinants for metallic ion resistance and enterotoxin B and C1 production were detected in heat-treated S. aureus 6344. Transformation employing chromosomal DNA containing the integrated plasmid resulted in excision and reestablishment of pZA10-related plasmids in the transformants. pZA10-linked resistance to cadmium, which was lost upon the integration of pZA10 into the host chromosome, reappeared in transformants carrying the excised plasmid.