Sequence variations in the introns of the triosephosphate isomerase genes of Oesophagostomum dentatum and O. quadrispinulatum

Abstract.

Degenerated primers were used to amplify DNA fragments of the triosephosphate isomerase (TPI) gene from complementary DNA (cDNA) and from genomic DNA of two species of porcine gastrointestinal nematodes, Oesophagostomum dentatum and O. quadrispinulatum. Polymerase chain reaction (PCR) fragments amplified from cDNA were 520 bp in size for both species, while genomic fragments were 1,035 bp for O. dentatum (GC-content: 45%) and 1,331 bp for O. quadrispinulatum (44%). Sequence analyses revealed blocks of high homology in the exons interrupted by more variable parts in the intron regions. Five exons were predicted from the genomic sequences in the conserved regions which corresponded to the respective cDNA sequences with 6% interspecific differences. The predicted protein sequences (161 amino acids) were 98% similar between the species and showed 71% similarity to the putative protein of Caenorhabditis elegans. As a housekeeping gene, TPI could be amplified from cDNA of both infectious third-stage larvae and adults. Interspecific variations in the non-coding regions allow the PCR-based differentiation of the two Oesophagostomum spp.