A panel of ten site-directed mutants of ε-toxin was generated. All of the mutated proteins expressed in were recognized in immunoblots by a neutralizing mAb raised against wild-type native ε-toxin. The cytotoxicity of the site-directed mutated toxins was assayed against MDCK cells. One mutation resulting in loss of activity in the assay was identified. This non-toxic protein was derived by substituting a proline for the histidine at residue 106 of the toxin. Immunization of mice with the non-toxic mutated ε-toxin resulted in the induction of a specific antibody response and immunized mice were protected against 1000 LD doses of wild-type recombinant ε-toxin.


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