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

Detalhes bibliográficos
Autor(a) principal: Gabriel, Jane Eyre
Data de Publicação: 2017
Outros Autores: de Souza, Emanuel Maltempi, de Carvalho, Francisco Allan Leandro, Madeira, Humberto Maciel França
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Biotemas (Online)
Texto Completo: https://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p1
Resumo: 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. 
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spelling Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbaginRegião predita de um promotor de genes induzíveis por paraquat da bactéria Chromobacterium violaceum sofre indução heteróloga pelo composto plumbaginThe 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. O objetivo deste estudo foi avaliar funcionalmente a influência do composto plumbagin sobre a indução heteróloga de uma região promotora predita de genes paraquat-induzíveis revelada durante as análises de anotação do genoma da bactéria Chromobacterium violaceum. A região promotora de interesse de C. violaceum foi amplificada a partir de sequências oligonucleotídicas específicas e clonada em vetor conjugativo, sendo acoplada à região codificante do gene lacZ de Escherichia coli. Em seguida, a indução heteróloga desse segmento regulatório foi estimada em cepa de E. coli na presença do plumbagin em uma concentração final de 50 µg/mL por mensurações dos níveis de expressão da enzima ?-galactosidase. Diferenças significativas nos níveis da ?-galactosidase foram observadas como resultado da ativação da região promotora de interesse pelo plumbagin na concentração testada em comparação às condições controle. Por outro lado, nenhum crescimento da cepa selvagem de C. violaceum foi observado durante a incubação dessas células em meio nutritivo contendo diferentes concentrações do plumbagin. As descobertas descritas aqui sugerem que uma região promotora predita para genes paraquat-induzíveis da bactéria C. violaceum sofra indução heteróloga pelo plumbagin, reforçando evidências acerca da caracterização funcional de motivos regulatórios intrínsecos a essa bactéria.Universidade Federal de Santa Catarina2017-05-25info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p110.5007/2175-7925.2017v30n2p1Biotemas; v. 30 n. 2 (2017); 1-62175-79250103-1643reponame:Biotemas (Online)instname:Universidade Federal de Santa Catarina (UFSC)instacron:UFSCenghttps://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p1/34112Copyright (c) 2017 Jane Eyre Gabriel, Emanuel Maltempi de Souza, Francisco Allan Leandro de Carvalho, Humberto Maciel França Madeirainfo:eu-repo/semantics/openAccessGabriel, Jane Eyrede Souza, Emanuel Maltempide Carvalho, Francisco Allan LeandroMadeira, Humberto Maciel França2019-03-07T08:53:41Zoai:periodicos.ufsc.br:article/45538Revistahttp://www.biotemas.ufsc.br/index.htmPUBhttps://periodicos.ufsc.br/index.php/biotemas/oai||carlospinto@ccb.ufsc.br2175-79250103-1643opendoar:2019-03-07T08:53:41Biotemas (Online) - Universidade Federal de Santa Catarina (UFSC)false
dc.title.none.fl_str_mv Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
Região predita de um promotor de genes induzíveis por paraquat da bactéria Chromobacterium violaceum sofre indução heteróloga pelo composto plumbagin
title Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
spellingShingle Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
Gabriel, Jane Eyre
title_short Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
title_full Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
title_fullStr Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
title_full_unstemmed Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
title_sort Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
author Gabriel, Jane Eyre
author_facet Gabriel, Jane Eyre
de Souza, Emanuel Maltempi
de Carvalho, Francisco Allan Leandro
Madeira, Humberto Maciel França
author_role author
author2 de Souza, Emanuel Maltempi
de Carvalho, Francisco Allan Leandro
Madeira, Humberto Maciel França
author2_role author
author
author
dc.contributor.author.fl_str_mv Gabriel, Jane Eyre
de Souza, Emanuel Maltempi
de Carvalho, Francisco Allan Leandro
Madeira, Humberto Maciel França
description 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. 
publishDate 2017
dc.date.none.fl_str_mv 2017-05-25
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p1
10.5007/2175-7925.2017v30n2p1
url https://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p1
identifier_str_mv 10.5007/2175-7925.2017v30n2p1
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://periodicos.ufsc.br/index.php/biotemas/article/view/2175-7925.2017v30n2p1/34112
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Universidade Federal de Santa Catarina
publisher.none.fl_str_mv Universidade Federal de Santa Catarina
dc.source.none.fl_str_mv Biotemas; v. 30 n. 2 (2017); 1-6
2175-7925
0103-1643
reponame:Biotemas (Online)
instname:Universidade Federal de Santa Catarina (UFSC)
instacron:UFSC
instname_str Universidade Federal de Santa Catarina (UFSC)
instacron_str UFSC
institution UFSC
reponame_str Biotemas (Online)
collection Biotemas (Online)
repository.name.fl_str_mv Biotemas (Online) - Universidade Federal de Santa Catarina (UFSC)
repository.mail.fl_str_mv ||carlospinto@ccb.ufsc.br
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