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Molecular cloning and characterization of a novel protein of Trichinella pseudospiralis excretory–secretory products

Published online by Cambridge University Press:  12 April 2024

I. Nagano ,
Z. Wu ,
T. Nakada ,
T. Boonmars  and
Y. Takahashi
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
T. Boonmars
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
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Abstract

A novel excretory–secretory (ES) protein of Trichinella pseudospiralis was produced. A cDNA library was constructed from mRNA of muscle larvae at 30 days post infection (p.i.) and immunoscreened with the antibody against ES products. A clone, designated Tp22-3, contained a cDNA transcript of 815 bp in length with a single open reading frame which encoded 244-amino acids (28407 Da in the estimated molecular mass). A database search revealed that no sequences had a homology to this predicted protein. The recombinant protein was produced in an Escherichia coli expression system. Stage specific expression of this protein was suggested from the following experiments. An antibody against the recombinant protein could stain proteins migrating at about 28 kDa (which is the expected size from the sequence) on Western blotting of crude extracts or ES products from 30 days p.i. muscle larvae, but failed to stain any proteins in crude extracts from newborn larvae or 15 days p.i. muscle larvae. The antibody reacted to the stichocytes of larvae at 30 days p.i., but did not react to 15 days p.i. muscle larvae. The production of an mRNA transcript for Tp22-3 gene was restricted largely to the 30 days p.i. muscle larvae and adult worms.

Type
Review Article
Information
Journal of Helminthology , Volume 76 , Issue 2 , June 2002 , pp. 165 - 170
Copyright
Copyright © Cambridge University Press 2002

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