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Cloning two full-length beta-tubulin isotype cDNAs from Cooperia oncophora, and screening for benzimidazole resistance-associated mutations in two isolates

Published online by Cambridge University Press:  05 December 2003

A. I. NJUE  and
R. K. PRICHARD
A. I. NJUE
Affiliation:
Institute of Parasitology, McGill University, 21 111 Lakeshore Road, Ste-Anne-de-Bellevue, Quebec, Canada H9X 3V9
R. K. PRICHARD
Affiliation:
Institute of Parasitology, McGill University, 21 111 Lakeshore Road, Ste-Anne-de-Bellevue, Quebec, Canada H9X 3V9
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Abstract

Two full-length beta-tubulin cDNAs, representing isotypes 1 and 2, were cloned from the cattle nematode Cooperia oncophora. The predicted protein sequences span 448 amino acids, and show a high degree of identity to beta-tubulins from other nematodes. While C. oncophora isotype 1 sequence had the highest identity to Haemonchus contortus isotype 1 and Teladorsagia circumcincta sequences (95% identity), the C. oncophora isotype 2 sequence was most similar to H. contortus isotype 2 and Trichostrongylus colubriformis (92% identity). Alignment of the two C. oncophora sequences with other trichostrongylid beta-tubulins deposited in GenBank showed a clear distinction between isotype 1 and 2 beta-tubulin classes. The two classes differed at 19 amino acid positions, most notably at the carboxy terminus. These isotype-defining residues were conserved among different trichostrongylid species within a class. Analysis of fragments of both genes revealed a high degree of genetic variability in coding and non-coding regions. However, all nucleotide differences detected in the coding region were silent, as they did not result in any amino acid substitution. Analysis of 2 groups of worms for the codon 200 polymorphism associated with benzimidazole resistance revealed a proportion of worms in 1 of the groups bearing a tyrosine at this position.

Keywords

Cooperia oncophorabeta-tubulincloningbenzimidazoleanthelmintic resistance
Type
Research Article
Information
Parasitology , Volume 127 , Issue 6 , December 2003 , pp. 579 - 588
Copyright
2003 Cambridge University Press

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