Molecular cloning and expression of the epidermolytic toxin A gene of Staphylococcus aureus

The gene coding for serotype A of epidermolytic (exfoliative) toxin has been cloned from Staphylococcus aureus in Escherichia coli phage lambda and plasmid vectors. The coding sequence for eta was localised by subcloning and transposon Tn5 mutagenesis experiments. The eta gene was probably expressed from its natural promoter in E. coli. The protein synthesised in E. coli was located predominantly in the periplasm. It was immunochemically indistinguishable from the toxin purified from S. aureus culture supernatants and had the same molecular weight. Furthermore, subcutaneous injection of this material caused epidermal splitting (the Nikolsky reaction) showing that it was biologically active. An eta shuttle plasmid was transformed into protoplasts of S. aureus. The level of expression of toxin in strain 8325-4 was shown to be dependent on the integrity of the agr gene which is known to be required for the expression of several exoproteins.