Molecular cloning and characterization of an IgE-reactive protein from Anisakis simplex: Ani s 1

Ingestion of the parasitic nematode Anisakis,simplex in undercooked fish ca n cause severe allergic reactions in some individuals. Using pooled human s era from sensitized patients we have probed an expression library for A. si mplex antigens. One positive clone was found to encode a full length 21 kDa protein with strong homology to nematode troponins. The recombinant protei n was expressed as a GST-fusion protein and found by immunoblot analysis to react with sera from 20% of allergic patients. The presence of functional EF-hand Ca2+ binding motifs was demonstrated by gel-shift analysis. (C) 200 0 Published by Elsevier Science B.V. All rights reserved.