Meloidogyne incognita: molecular cloning and characterization of a cDNA encoding a cathepsin D-like aspartic proteinase

• Autor(a) principal: Fragoso, Rodrigo da Rocha
• Data de Publicação: 2009
• Outros Autores: Lourenço, Isabela Tristan, Batista, João Aguiar Nogueira, Oliveira-Neto, Osmundo Brilhante, Silva, Maria Cristina Mattar, Rocha, Thales Lima, Coutinho, Marise Ventura, Grossi-de-Sa, Maria Fátima
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UCB
• Texto Completo: http://twingo.ucb.br:8080/jspui/handle/10869/482; https://repositorio.ucb.br:9443/jspui/handle/123456789/7686
• topic: cDNA cloning; Meloidogyne; Nematode; Aspartic proteinase; Cathepsin D; RKM-Root-knot nematode; CN-cyst nematode; L2 - second-stage larva or juvenile; AP- aspartic proteinase; RACE- rapid amplification; EST, expressed sequence tag; RT-PCR- reverse transcription and polymerase chain reaction; ORF- open read frame

Herein we describe the cloning and characterization of a cDNA encoding an aspartic proteinase from the root-knot nematode Meloidogyne incognita. Using PCR techniques, a 1471-bp cDNA fragment encoding a cathepsin D-like (Mi-asp1) transcript was isolated from second- stage larvae mRNA. Its predicted amino acid sequence comprises a pro-region of 71 amino acid residues and a mature protease of 378 amino acid residues with a predicted molecular mass of 41.502 kDa. Protein sequence comparisons of Mi-asp1 with GenBank ™ (DQ360827) sequences showed 59–71% identity with nematode- specific cathepsin D-like aspartic proteinases. Southern blot analysis, RT-PCR amplification and EST mining suggest the existence of a developmentally expressed gene family encoding aspartic proteinases in M. incognita. Mi-asp1 may represent a potential target for molecular intervention for the purposes of plant–parasitic nematode control.