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Provedor de dados:  Genet. Mol. Biol.
País:  Brazil
Título:  Expression, purification and DNA-binding activities of two putative ModE proteins of Herbaspirillum seropedicae (Burkholderiales, Oxalobacteraceae)
Autores:  Souza,André L.F.
Chubatsu,Leda S.
Souza,Emanuel M.
Pedrosa,Fábio O.
Monteiro,Rose A.
Rego,Fabiane G.M.
Rigo,Liu U.
Data:  2008-01-01
Ano:  2008
Palavras-chave:  Herbaspirillum seropedicae
ModE1 protein
ModE2 protein
Resumo:  In prokaryotes molybdenum is taken up by a high-affinity ABC-type transporter system encoded by the modABC genes. The endophyte β-Proteobacterium Herbaspirillum seropedicae has two modABC gene clusters and two genes encoding putative Mo-dependent regulator proteins (ModE1 and ModE2). Analysis of the amino acid sequence of the ModE1 protein of H. seropedicae revealed the presence of an N-terminal domain containing a DNA-binding helix-turn-helix motif (HTH) and a C-terminal domain with a molybdate-binding motif. The second putative regulator protein, ModE2, contains only the helix-turn-helix motif, similar to that observed in some sequenced genomes. We cloned the modE1 (810 bp) and modE2 (372 bp) genes and expressed them in Escherichia coli as His-tagged fusion proteins, which we subsequently purified. The over-expressed recombinant His-ModE1 was insoluble and was purified after solubilization with urea and then on-column refolded during affinity chromatography. The His-ModE2 was expressed as a soluble protein and purified by affinity chromatography. These purified proteins were analyzed by DNA band-shift assays using the modA2 promoter region as probe. Our results indicate that His-ModE1 and His-ModE2 are able to bind to the modA2 promoter region, suggesting that both proteins may play a role in the regulation of molybdenum uptake and metabolism in H. seropedicae.
Tipo:  Info:eu-repo/semantics/article
Idioma:  Inglês
Editor:  Sociedade Brasileira de Genética
Relação:  10.1590/S1415-47572008000400022
Formato:  text/html
Fonte:  Genetics and Molecular Biology v.31 n.3 2008
Direitos:  info:eu-repo/semantics/openAccess

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