A novel cell wall degrading enzyme was identi¿ ed as a part of cDNA-AFLP project that compared gene expression pro¿ les in ¿ ve different stages of Globodera rostochiensis. One Transcript Derived Fragment (TDF), predominantly expressed in infective second stage juveniles, encoded a partial open reading frame showing high similarity to bacterial and fungal pectate lyases from the polysaccharide lyase 3 family (EC 188.8.131.52). The corresponding full-length cDNA designated Gr-Pel2 harbours an open reading frame of 759 bp, encoding polypeptide with molecular mass 27 kDa. Gr-Pel2 is signi¿ cantly different from previously identi¿ ed Gr- Pel1 (30% identities and 46% similarities). Sequence was predicted to have signal peptide for secretion at its N-terminal end. Digoxigenin-labelled DNA probe hybridised speci¿ cally to the subventral pharyngeal glands, which are responsible for secreting all currently known cell wall degrading enzymes. Heterologous expression of Gr-Pel2 in Pichia pastoris is in progress, in order to assess its biochemical properties. Identi¿ cation of another pectate lyase gene shows the complexity of the pectinolytic system in nematode secretions.
|Title of host publication
|Nematology Symposium abstracts
|Place of Publication
|Brill Academic Publishers
|Published - 2002