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Molecular cloning and characterization of Echinostoma caproni heat shock protein-70 and differential expression in the parasite derived from low- and high-compatible hosts

Published online by Cambridge University Press:  28 August 2008

M. HIGÓN ,
C. MONTEAGUDO ,
B. FRIED ,
J. G. ESTEBAN ,
R. TOLEDO  and
A. MARCILLA
M. HIGÓN
Affiliation:
Departamento de Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain
C. MONTEAGUDO
Affiliation:
Departamento de Patología, Facultat de Medicina, Av. Blasco Ibañez, 17, Valencia, Spain
B. FRIED
Affiliation:
Department of Biology, Lafayette College, Easton, Pennsylvania 18042, USA
J. G. ESTEBAN
Affiliation:
Departamento de Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain
R. TOLEDO
Affiliation:
Departamento de Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain
A. MARCILLA*
Affiliation:
Departamento de Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain
*
*Corresponding author: Departamento de Biología Celular y Parasitología, Facultat de Farmàcia, Universitat de Valencia, Av. V.A. Estellés, s/n, 46100 Burjassot, Valencia, Spain. Tel: +34 963544491. Fax: +34 963544769. E-mail: [email protected]
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Summary

We cloned and expressed Echinostoma caproni HSP70 in Escherichia coli. This molecule presents an open reading frame (ORF) of 655 amino acids, and a theoretical molecular weight of 71 kDa. E. caproni HSP70 protein showed a high homology to other helminth molecules, major differences being located in the C-terminal region of the molecule, with a hydrophobic portion. Studies of protein and messenger RNA (mRNA) expression revealed a distinct pattern, depending on the host (low- or high-compatible). Specific polyclonal antisera raised against the recombinant protein expressed in Escherichia coli demonstrated its selective presence in excretory/secretory products (ESP) of adult parasites obtained from high-compatible hosts. Immunological studies showed clearly the association of HSP70 with the parasite surface and other structures, including eggs.

Keywords

Echinostoma caproniheat shock proteinHSP70molecular cloning and expressionimmunolocalizationparasitetrematode
Type
Original Articles
Information
Parasitology , Volume 135 , Issue 12 , October 2008 , pp. 1469 - 1477
Copyright
Copyright © 2008 Cambridge University Press

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