skip to main content
Idioma:

Isolation of Xanthomonas axonopodis pv citri knockout mutants affected in the oppA gene encoding the oligopeptide uptake system

E. E. Oshiro R. S. L Nepomuceno; C. F Suzuki; W Kruger; J. B Faria; Rita de Cássia Café Ferreira; Luís Carlos de Souza Ferreira; Sociedade Brasileira de Bioquímica e Biologia Molecular - SBBq (33. 2004 Caxambu)

Programa e resumos Caxambu, MG

Caxambu 2004

Item não circula. Consulte sua biblioteca.(Acessar)

  • Título:
    Isolation of Xanthomonas axonopodis pv citri knockout mutants affected in the oppA gene encoding the oligopeptide uptake system
  • Autor: E. E. Oshiro
  • R. S. L Nepomuceno; C. F Suzuki; W Kruger; J. B Faria; Rita de Cássia Café Ferreira; Luís Carlos de Souza Ferreira; Sociedade Brasileira de Bioquímica e Biologia Molecular - SBBq (33. 2004 Caxambu)
  • Assuntos: MICROBIOLOGIA
  • É parte de: Programa e resumos Caxambu, MG
  • Notas Locais: Disponível somente em CD-ROM
  • Descrição: The eubacteria Xanthomonas axonopodis pv citri (Xac) is the causative agent of the citrus canker, which causes signi_cant economic losses to farmers and the citrus industry. Identi_cation of genes involved in virulence mechanisms or encoding possible targets for therapeutic control represent a priority for physiological studies of this microorganism. As a step toward the evaluation of genes involved in the nutritional strategies of Xanthomonas species, we describe the isolation of knockout mutants a_ected in the gene encoding the oligopeptide binding protein (OppA), the periplasmic soluble component of the oligopeptide uptake system (Opp) found in the Xac genome but not in other Xanthomonas species. A knockout copy of the oppA gene was generated by cloning a spectinomycin resistance cassette into the unique EcoRV (Biolabs) site located in the middle region of the cloned gene. The mutated oppAgene was sub-cloned in the R6K-derived suicide plasmid pWR-SS. The resulting recombinant plasmid was introduced into the Xac 306 strain by electroporation using optimal conditions previously established by our group. Colonies were selected for allelic-replacement after a two-step procedure based _rst on resistance to spectinomycin and, in a second step, by resistance to 5 per cent sucrose due to loss of the sacB gene present in the suicide vector. Up to 22,85 per cent (8/35) of the spectinomycin-resistant and sucrose-resistant colonies were shown to carry the
    mutated knockout allele as a result of a two-cross over event, as demonstrated by Southern blots and PCR analyses. Further experiments were carried out to evaluate the role of the oppA-de_cient mutants on the resistance to several antibiotics and ability to grow on media supplemented with peptides as nutrient sources. Such results contribute for a better understanding of the Opp system on the physiology and nutritional strategies of Xac
  • Editor: Caxambu
  • Data de criação/publicação: 2004
  • Formato: res. F54.
  • Idioma: Inglês
Links
Voltar para lista de resultados
Ir para página anteriorAnterior Resultado  14 AvançarIr para próxima página

  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

Buscando em bases de dados remotas. Favor aguardar.