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Cloning and characterization of a β-galactoside-binding protein (galectin) from the gut of the gastrointestinal nematode parasite Haemonchus contortus

  • G. F. J. NEWLANDS (a1), P. J. SKUCE (a1), D. P. KNOX (a1), S. K. SMITH (a1) and W. D. SMITH (a1)...

Abstract

A cDNA encoding a β-galactoside-binding lectin (galectin) was identified by immunoscreening a Haemonchus contortus cDNA library with antisera from lambs vaccinated with a membrane protein complex (H-gal-GP) derived from the parasites' gut. The cDNA sequence, exhibiting a tandem repeat structure and designated Hco-gal-2, showed significant levels of similarity with galectins from several species of nematode as well as mammalian galectin type 4. Native galectin was preferentially extracted from the H-gal-GP complex and also from an insoluble membrane fraction prepared from adult worms using lactose–agarose affinity chromatography. The affinity-purified material had apparent molecular mass of around 35 kDa with 3 distinct bands visible on SDS–PAGE. All 3 bands were identified as galectins by reaction with antiserum raised to recombinant Hco-GAL-2 on Western blot. To determine whether H. contortus galectins have any protective capacity against infection lambs were vaccinated with affinity-purified galectin and subsequently given a single challenge infection. Vaccination did not confer any protection against infection with H. contortus as judged by faecal egg output or worm counts.

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Corresponding author

Corresponding author: Tel: +44 (0)131 445 5111. Fax: +44 (0)131 445 6111. E-mail: newlg@mri.sari.ac.uk

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Nucleotide and amino acid sequence data reported in this paper are available in the GenBank database under the accession number AF036098.

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Keywords

Cloning and characterization of a β-galactoside-binding protein (galectin) from the gut of the gastrointestinal nematode parasite Haemonchus contortus

  • G. F. J. NEWLANDS (a1), P. J. SKUCE (a1), D. P. KNOX (a1), S. K. SMITH (a1) and W. D. SMITH (a1)...

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