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Molecular cloning and characterization of an Rcd1-like protein in excretory-secretory products of Trichinella pseudospiralis

Published online by Cambridge University Press:  10 August 2006

I. NAGANO ,
Z. WU  and
Y. TAKAHASHI
I. NAGANO
Affiliation:
Department of Parasitology, Gifu University Graduate School of Medicine, Yanagido 1-1 Gifu, 501-1194 Japan
Z. WU
Affiliation:
Department of Parasitology, Gifu University Graduate School of Medicine, Yanagido 1-1 Gifu, 501-1194 Japan
Y. TAKAHASHI
Affiliation:
Department of Parasitology, Gifu University Graduate School of Medicine, Yanagido 1-1 Gifu, 501-1194 Japan
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Abstract

A cDNA library was constructed from muscle larvae of Trichinella pseudospiralis. A cDNA clone, designated as Tp8 contained a cDNA transcript of 1326 bp length with a single open reading frame, which encoded 303 amino acid residues (34187 Da, estimated molecular mass). The predicted amino acid sequence of the clone had an identity of approximately 60% to the Rcd1 (Required cell differentiation 1) -like proteins among a wide range of organisms. Real-time quantitative polymerase chain reaction results showed that the transcription level of Tp8 gene reached the highest value in adult worms, and that the transcription level in muscle larvae before stichosome formation was higher than in muscle larvae after stichosome formation. The recombinant Tp8 protein migrated at 37 kDa and reacted to antibody against T. pseudospiralis excretory-secretory (E-S) products and sera from mice infected with T. pseudospiralis. An antibody against the Tp8 recombinant protein could stain proteins migrating at approximately 34 kDa (which is the expected size from the sequence) on Western blotting of E-S products from muscle larvae. An immunocytochemical study showed that the Tp8 protein was present within the stichocyte of muscle larvae and adults worms.

Keywords

Trichinella pseudospiralisRcd1molecular cloningexcretory-secretory products
Type
Research Article
Information
Parasitology , Volume 133 , Issue 6 , December 2006 , pp. 785 - 792
Copyright
2006 Cambridge University Press

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