Isolation and localisation of an annexin gene (gp-nex) from the potato cyst nematode, Globodera pallida
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Abstract
A full length cDNA clone was isolated by screening a mixed stage Globodera pallida expression library using the monospecific polyclonal antibody IACR-PC320. Sequence comparison indicated that the predicted 332 amino acid protein encoded by the cDNA was a member of the annexin gene family named gp-nex and very similar to annexins found in other nematode species. Annexins are calcium-dependent phospholipid binding proteins characterised by four repeated domains approximately 70 amino acids in length. The predicted amino acid sequence of the protein did not contain an N-terminal secretion signal peptide; however, the protein was shown to be present in excretory/secretory products from G. pallida second stage juveniles treated with the neurotransmitter 5 methoxy-N, N dimethyl tryptamine. Gp-nex encodes a protein of 35 kDa and was immunolocalised in the amphids, genital primordium and in the constraining muscles above and below the metacorpus pump chamber of G. pallida second stage juveniles. It is also present in eggs and adult females of the two species of the potato cyst nematodes.