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Molecular cloning and characterization of a 21 kDa protein secreted from Trichinella pseudospiralis

Published online by Cambridge University Press:  22 February 2007

I. Nagano
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
Z. Wu
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
T. Nakada
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
A. Matsuo
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
Y. Takahashi*
Affiliation:
Department of Parasitology, Gifu University School of Medicine, Tsukasa 40, Gifu 500-8705, Japan
*
*Author for correspondence Fax: 058 267 2960 E-mail: yu3@cc.gifu-u.ac.jp

Abstract

Recombinant protein was produced from the cDNA library of Trichinella pseudospiralis, which seemed to form part of the excretory–secretory (ES) products. The library was constructed from cDNA of muscle larvae at 1 month post-infection, and immunoscreened with antibody against T. pseudospiralis ES products. A clone, designated Tp21-3, contained a cDNA transcript of 657 bp in length with a single open reading frame, which encoded 172 amino acids (19617 Da in the estimated molecular mass). The predicted amino acid sequence of clone Tp21-3 had a similarity of 76% to that of clone ORF 17.20 (GenBank under accession number U88239) from T. spiralis. The recombinant fusion proteins encoded by clone Tp21-3 were produced in an Escherichia coli expression system and affinity purified. On Western blotting analysis, Tp21-3 recombinant proteins migrated at 40 kDa and reacted to antibody against T. pseudospiralis ES products and T. pseudospiralis-infected sera. Sera were developed against Tp 21-3 recombinant proteins, which reacted to a single band migrating at 21 kDa in crude worm extract and ES products from T. pseudospiralis on Western blotting analysis, and reacted with stichocytes of T. pseudospiralis on immunohistochemical staining.

Type
Research Article
Copyright
Cambridge University Press 2001

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Molecular cloning and characterization of a 21 kDa protein secreted from Trichinella pseudospiralis
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