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Coat proteins from mature spores of two enterotoxin-positive (Ent+) and two enterotoxin-negative (Ent+) strains of Clostridium perfringens were solubilized using 50 mm-dithiothreitol and 1% sodium dodecyl sulphate at pH 9·7, and alkylated using 110 mm-iodoacetamide to prevent aggregation. The coat proteins and C. perfringens type A enterotoxin (CPE) were separated by SDS-PAGE and analysed by Western blotting using anti-CPE antibody. As previously reported, CPE aggregated in the presence of SDS, but no aggregation occurred at concentrations below 15 μg CPE ml−. Two CPE-related proteins (34 and 48 kDa) were found in the solubilized spore coat protein of Ent+ strains while only the 48 kDa CPE-related protein was found in the spore coat fraction of Ent+ strains. CPE-related proteins comprised 2·7% and 0·8% of the total solubilized coat protein of Ent+ and Ent+ strains respectively. CPE-related proteins could be extracted from the spores with 1 % SDS alone. They could also be released by disruption of whole spores, indicating that the CPE-related proteins may be in the spore core or trapped between the core and coat layers. The results suggest that CPE is not a major structural component of the coat fraction of C. perfringens spores.
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