Isolation and characterization of a cDNA encoding a serine proteinase from the root-knot nematode meloidogyne incognita

• Autor(a) principal: Fragoso, Rodrigo da Rocha
• Data de Publicação: 2005
• Outros Autores: Batista, João Aguiar Nogueira, Oliveira Neto, Osmundo Brilhante, Grossi- de-Sá, Maria Fátima
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UCB
• Texto Completo: http://twingo.ucb.br:8080/jspui/handle/10869/435; https://repositorio.ucb.br:9443/jspui/handle/123456789/7645
• topic: cDNA cloning; Meloidogyne; Nematode; Serine proteinase

This report describes the Wrst serine proteinase gene isolated from the sedentary nematode Meloidogyne incognita. Using degenerate primers, a 1372 bp cDNA encoding a chymotrypsin-like serine proteinase (Mi-ser1) was ampliWed from total RNA of adult females by RT-PCR and 5_ and 3_ rapid ampliWcation of cDNA ends. The deduced amino acid sequence of Mi-ser1 encoded a putative signal peptide and a prodomain of 22 and 33 amino acids, respectively, and a mature proteinase of 341 amino acids with a predicted molecular mass of 37,680Da. Sequence identity with the top serine proteinases matches from the databases ranged from 23 to 27%, including sequences from insects, mammals, and other nematodes. Southern blot analysis suggested that Mi-ser1 is encoded by a single or few gene copies. The pattern of evelopmental expression analyzed by Northern blot and RT-PCR indicated that Mi-ser1was transcribed mainly in females. The domain architecture composed of a single chymotrypsin-like catalytic domain and the detection of a putative signal peptide suggested a digestive role for Mi-ser1.