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The complete DNA sequence of a cloned alpha-toxin gene from Staphylococcus aureus was determined. The amino acid sequence of the alpha-toxin protein, predicted from the DNA sequence, was described and compared with published data. The primary product of the cloned alpha-toxin gene contained a 26-amino-acid leader sequence which possessed characteristic features of a signal sequence involved in secretion. The mature alpha-toxin protein had a molecular size of 33,000 and contained only three short regions of high hydrophobicity in addition to a number of short, weakly hydrophobic regions.