Heterologous expression of predicted promoter site for paraquat-inducible genes of the bacterium Chromobacterium violaceum is increased by plumbagin
Autor(a) principal: | |
---|---|
Data de Publicação: | 2017 |
Outros Autores: | , , |
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. |
id |
UFSC-14_527a61ccff03395e5a8055f812794b1c |
---|---|
oai_identifier_str |
oai:periodicos.ufsc.br:article/45538 |
network_acronym_str |
UFSC-14 |
network_name_str |
Biotemas (Online) |
repository_id_str |
|
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 |
_version_ |
1799940554013802496 |