Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin

Authors

  • Jane Eyre Gabriel Universidade Federal do Vale do São Francisco UNIVASF
  • Emanuel Maltempi de Souza Universidade Federal do Paraná UFPR
  • Francisco Allan Leandro de Carvalho Universidade Federal do Vale do São Francisco UNIVASF
  • Humberto Maciel França Madeira Pontifícia Universidade Católica do Paraná PUCPR

DOI:

https://doi.org/10.5007/2175-7925.2017v30n2p1

Abstract

The aim of this study was to evaluate functionally the effect of plumbagin on the heterologous expression of a predicted promoter region of open reading frames of paraquat-inducible (pqi) genes revealed during genome annotation analyses of the bacterium Chromobacterium violaceum. First, the promoter of interest was amplified using specific primers and cloned into a conjugative vector carrying the Escherichia coli lacZ gene without a promoter. The heterologous expression of the predicted promoter region was then examined in the presence of 50 µg/mL plumbagin by ?-galactosidase expression assays. Significant differences were detected in the levels of ?-galactosidase as a result of the activation of the promoter region of interest in response to plumbagin at the concentration tested. On the other hand, no growth of the wild strain of C. violaceum was found during its incubation in nutrient broth medium containing different concentrations of plumbagin compared to control group. The findings described herein demonstrate that the heterologous expression of a predicted promoter site of pqi genes of C. violaceum is induced by plumbagin in a fusion strain, giving insights into the functional characterization of intrinsic regulatory DNA motifs annotated in this bacterial genome.

 

Author Biographies

Jane Eyre Gabriel, Universidade Federal do Vale do São Francisco UNIVASF

Graduação em Ciências Biológicas pelo Instituto de Biociências, Letras e Ciências Exatas (1989), mestrado em Zootecnia pela Universidade Estadual Paulista Júlio de Mesquita Filho (1996) e doutorado em Ciências (Energia Nuclear na Agricultura) pela Universidade de São Paulo (2001). Tem experiência na área de Biologia Molecular Animal, Bioinformática, atuando principalmente nos seguintes temas: expressão gênica animal, genômica e proteômica, histologia geral.

Emanuel Maltempi de Souza, Universidade Federal do Paraná UFPR

Graduação em Farmacia e Bioquimica pela Universidade Federal do Paraná (1984) e doutorado em Ciências (Bioquímica) pela Universidade Federal do Paraná (1990). Tem experiência na área de Bioquímica, com ênfase em Biologia Molecular. Atua principalmente nos seguintes temas: fixação biológica de nitrogênio, regulação da expressão gênica em Azospirillum brasilense e Herbaspirillium seropedicae, expressão e purificação de proteínas em E. coli e mecanismos moleculares da interacao planta-bactéria.

 

Francisco Allan Leandro de Carvalho, Universidade Federal do Vale do São Francisco UNIVASF

Graduação em Tecnologia em Alimentos pelo Instituto Federal do Sertão Pernambucano (2008) e mestrado em Ciência Animal pela Universidade Federal do Vale do São Francisco (2013). Atualmente é doutorando em Engenharia de Alimentos pela Universidade de São Paulo FZEA-USP e servidor (Técnico de Nível Superior) da Universidade Federal do Vale do São Francisco.

Humberto Maciel França Madeira, Pontifícia Universidade Católica do Paraná PUCPR

Graduação em Engenharia Agronômica pela Universidade de São Paulo (USP) (1988), mestrado em Nutrição Animal pela USP (1992) e Ph.D. em Animal Science - University of Nebraska (1998). Possui estágio pós-doutoral pela USP (1998-1999) e pela University of Illinois at Urbana-Champaign (2012). Tem experiência na área de Biologia Molecular e Microbiologia, com ênfase em Microbiologia Gastrintestinal, atuando principalmente nos seguintes temas: microbiologia de anaeróbios, genômica, bioinformática, biotecnologia, antimicrobianos, nutrição de ruminantes.

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Published

2017-05-25

Issue

Vol. 30 No. 2 (2017)

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Artigos

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