Evaluation of reference genes for insect olfaction studies

Detalhes bibliográficos
Autor(a) principal: Omondi, Bonaventure Aman
Data de Publicação: 2015
Outros Autores: Estivalis, José Manuel Latorre, Oliveira, Ivana Helena Rocha, Ignell, Rickard, Lorenzo, Marcelo Gustavo
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/12669
Resumo: Department of Plant Protection Biology. Chemical Ecology Unit. SLU, Alnarp, Sweden / Bioversity International. Consultative Group for International Agricultural Research. Bujumbura, Burundi.
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spelling Omondi, Bonaventure AmanEstivalis, José Manuel LatorreOliveira, Ivana Helena RochaIgnell, RickardLorenzo, Marcelo Gustavo2016-02-01T17:35:45Z2016-02-01T17:35:45Z2015OMONDI, Bonaventure Aman et al. Evaluation of reference genes for insect olfaction studies. Parasit Vectors, v. 8, p. 1-15, 2015.1756-3305https://www.arca.fiocruz.br/handle/icict/1266910.1186/s13071-015-0862-x.engBioMed CentralEvaluation of reference genes for insect olfaction studiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleDepartment of Plant Protection Biology. Chemical Ecology Unit. SLU, Alnarp, Sweden / Bioversity International. Consultative Group for International Agricultural Research. Bujumbura, Burundi.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Comportamento do Vetor e Grupo de Interações com Patogenos. Belo Horizonte, MG, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Comportamento do Vetor e Grupo de Interações com Patógenos. Belo Horizonte, MG, Brasil.Department of Plant Protection Biology. Chemical Ecology Unit. SLU, Alnarp, Sweden.Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Comportamento de Vetores e Interação com Patógenos. Belo Horizonte, MG, Brasil.Background: Quantitative reverse transcription PCR (qRT-PCR) is a robust and accessible method to assay gene expression and to infer gene regulation. Being a chain of procedures, this technique is subject to systematic error due to biological and technical limitations mainly set by the starting material and downstream procedures. Thus, rigorous data normalization is critical to grant reliability and repeatability of gene expression quantification by qRT-PCR. A number of ‘housekeeping genes’, involved in basic cellular functions, have been commonly used as internal controls for this normalization process. However, these genes could themselves be regulated and must therefore be tested a priori. Methods: We evaluated eight potential reference genes for their stability as internal controls for RT-qPCR studies of olfactory gene expression in the antennae of Rhodnius prolixus, a Chagas disease vector. The set of genes included were: α-tubulin; β-actin; Glyceraldehyde-3-phosphate dehydrogenase; Eukaryotic initiation factor 1A; Glutathione-S-transferase; Serine protease; Succinate dehydrogenase; and Glucose-6-phosphate dehydrogenase. Five experimental conditions, including changes in age,developmental stage and feeding status were tested in both sexes. Results: We show that the evaluation of candidate reference genes is necessary for each combination of sex, tissue and physiological condition analyzed in order to avoid inconsistent results and conclusions. Although, Normfinder and geNorm software yielded different results between males and females, five genes (SDH, Tub, GAPDH, Act and G6PDH) appeared in the first positions in all rankings obtained. By using gene expression data of a single olfactory coreceptor gene as an example, we demonstrated the extent of changes expected using different internal standards. Conclusions; This work underlines the need for a rigorous selection of internal standards to grant the reliability of normalization processes in qRT-PCR studies. Furthermore, we show that particular physiological or developmental conditions require independent evaluation of a diverse set of potential reference genes.RT-qPCRReference genesNormalization processOlfaction and triatominesinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/12669/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALEvaluation of reference genes for insect olfaction studies.pdfEvaluation of reference genes for insect olfaction studies.pdfapplication/pdf13551079https://www.arca.fiocruz.br/bitstream/icict/12669/2/Evaluation%20of%20reference%20genes%20for%20insect%20olfaction%20studies.pdfe23c2a48fc3b69cadff7f0386fc1d505MD52TEXTEvaluation of reference genes for insect olfaction studies.pdf.txtEvaluation of reference genes for insect olfaction studies.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv Evaluation of reference genes for insect olfaction studies
title Evaluation of reference genes for insect olfaction studies
spellingShingle Evaluation of reference genes for insect olfaction studies
Omondi, Bonaventure Aman
RT-qPCR
Reference genes
Normalization process
Olfaction and triatomines
title_short Evaluation of reference genes for insect olfaction studies
title_full Evaluation of reference genes for insect olfaction studies
title_fullStr Evaluation of reference genes for insect olfaction studies
title_full_unstemmed Evaluation of reference genes for insect olfaction studies
title_sort Evaluation of reference genes for insect olfaction studies
author Omondi, Bonaventure Aman
author_facet Omondi, Bonaventure Aman
Estivalis, José Manuel Latorre
Oliveira, Ivana Helena Rocha
Ignell, Rickard
Lorenzo, Marcelo Gustavo
author_role author
author2 Estivalis, José Manuel Latorre
Oliveira, Ivana Helena Rocha
Ignell, Rickard
Lorenzo, Marcelo Gustavo
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Omondi, Bonaventure Aman
Estivalis, José Manuel Latorre
Oliveira, Ivana Helena Rocha
Ignell, Rickard
Lorenzo, Marcelo Gustavo
dc.subject.en.pt_BR.fl_str_mv RT-qPCR
Reference genes
Normalization process
Olfaction and triatomines
topic RT-qPCR
Reference genes
Normalization process
Olfaction and triatomines
description Department of Plant Protection Biology. Chemical Ecology Unit. SLU, Alnarp, Sweden / Bioversity International. Consultative Group for International Agricultural Research. Bujumbura, Burundi.
publishDate 2015
dc.date.issued.fl_str_mv 2015
dc.date.accessioned.fl_str_mv 2016-02-01T17:35:45Z
dc.date.available.fl_str_mv 2016-02-01T17:35:45Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv OMONDI, Bonaventure Aman et al. Evaluation of reference genes for insect olfaction studies. Parasit Vectors, v. 8, p. 1-15, 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/12669
dc.identifier.issn.pt_BR.fl_str_mv 1756-3305
dc.identifier.doi.none.fl_str_mv 10.1186/s13071-015-0862-x.
identifier_str_mv OMONDI, Bonaventure Aman et al. Evaluation of reference genes for insect olfaction studies. Parasit Vectors, v. 8, p. 1-15, 2015.
1756-3305
10.1186/s13071-015-0862-x.
url https://www.arca.fiocruz.br/handle/icict/12669
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/12669/1/license.txt
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