Data/hora: 09/06/2024 02:25:49

Biblioteca(s): | Embrapa Agrobiologia.
Data corrente: | 22/04/2008
Data da última atualização: | 16/03/2015
Autoria: | ROUWS, L. F. M.; SIMÕES ARAÚJO, J. L.; HEMERLY, A. S.; BALDANI, J. I.
Afiliação: | Luc F. M. Rouws, UFRRJ; Jean Luis Simões Araújo, Embrapa Agrobiologia; Adriana S. Hemerly, Instituto de Pesquisas do Jardim Botânico do RJ; José Ivo Baladani, Embrapa Agrobiologia.
Título: | Validation of a Tn5 transposon mutagenesis system for Gluconacetobacter diazotrophicus through characterization of a flagellar mutant.
Ano de publicação: | 2008
Fonte/Imprenta: | Archives of Microbiology, New York, v. 189, n. 4, p. 397-405, apr. 2008.
Idioma: | Inglês
Notas: | Parceria: UFRRJ; Instituto de Pesquisas do Jardim Botânico do Rio de Janeiro.
Conteúdo: | Gluconacetobacter diazotrophicus is a nitrogen-fixing bacterium, which was originally isolated from the interior of sugarcane plants. The genome of strain PAL5 of G. diazotrophicus has been completely sequenced and a next step is the functional characterization of its genes. The aim of this study was to establish an efficient mutagenesis method, using the commercial Tn5 transposon EZ::Tn5?<KAN-2>Tnp Transposome? (Epicentre). Up to 1 × 106 mutants per microgram of transposome were generated in a single electroporation experiment. Insertion-site flanking sequences were amplified by inverse PCR and sequenced for 31 mutants. For ten of these mutants, both insertion flanks could be identified, confirming the 9 bp duplication that is typical for Tn5 transposition. Insertions occurred in a random fashion and were genetically stable for at least 50 generations. One mutant had an insertion in a homolog of the flagellar gene flgA, and was therefore predicted to be affected in flagella-dependent traits and used to validate the applied mutagenesis methodology. This mutant lacked flagella and was non-motile on soft agar. Interestingly, it was also strongly affected in the ability to form biofilm on glass wool.
Palavras-Chave: | Bactéria endofítica.
Thesaurus Nal: | Gluconacetobacter diazotrophicus.
Categoria do assunto: | --
Registro original: | Embrapa Agrobiologia (CNPAB)
Exemplares: | CNPAB - (35051-1) - UPC - AP - -- -  -  - 0024125 - DIS