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

Published online by Cambridge University Press:  01 November 1999

G. F. J. NEWLANDS
Affiliation:
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 0PZ, UK
P. J. SKUCE
Affiliation:
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 0PZ, UK
D. P. KNOX
Affiliation:
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 0PZ, UK
S. K. SMITH
Affiliation:
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 0PZ, UK
W. D. SMITH
Affiliation:
Moredun Research Institute, International Research Centre, Pentland Science Park, Bush Loan, Penicuik EH26 0PZ, UK

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.

Type
Research Article
Copyright
1999 Cambridge University Press

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Footnotes

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