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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
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: antonio.marcilla@uv.es

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.

Type
Original Articles
Copyright
Copyright © 2008 Cambridge University Press

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