Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Microbiology |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259 |
Resumo: | Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis. |
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Brazilian Journal of Microbiology |
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Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidisPandora neoaphidisReference genesQuantitative PCRAbstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis.Sociedade Brasileira de Microbiologia2016-03-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259Brazilian Journal of Microbiology v.47 n.1 2016reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1016/j.bjm.2015.11.031info:eu-repo/semantics/openAccessChen,ChunXie,TingnaYe,SudanJensen,Annette BruunEilenberg,J ørgeneng2016-03-01T00:00:00Zoai:scielo:S1517-83822016000100259Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2016-03-01T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false |
dc.title.none.fl_str_mv |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
title |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
spellingShingle |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis Chen,Chun Pandora neoaphidis Reference genes Quantitative PCR |
title_short |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
title_full |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
title_fullStr |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
title_full_unstemmed |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
title_sort |
Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis |
author |
Chen,Chun |
author_facet |
Chen,Chun Xie,Tingna Ye,Sudan Jensen,Annette Bruun Eilenberg,J ørgen |
author_role |
author |
author2 |
Xie,Tingna Ye,Sudan Jensen,Annette Bruun Eilenberg,J ørgen |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Chen,Chun Xie,Tingna Ye,Sudan Jensen,Annette Bruun Eilenberg,J ørgen |
dc.subject.por.fl_str_mv |
Pandora neoaphidis Reference genes Quantitative PCR |
topic |
Pandora neoaphidis Reference genes Quantitative PCR |
description |
Abstract The selection of suitable reference genes is crucial for accurate quantification of gene expression and can add to our understanding of host–pathogen interactions. To identify suitable reference genes in Pandora neoaphidis, an obligate aphid pathogenic fungus, the expression of three traditional candidate genes including 18S rRNA(18S), 28S rRNA(28S) and elongation factor 1 alpha-like protein (EF1), were measured by quantitative polymerase chain reaction at different developmental stages (conidia, conidia with germ tubes, short hyphae and elongated hyphae), and under different nutritional conditions. We calculated the expression stability of candidate reference genes using four algorithms including geNorm, NormFinder, BestKeeper and Delta Ct. The analysis results revealed that the comprehensive ranking of candidate reference genes from the most stable to the least stable was 18S (1.189), 28S (1.414) and EF1 (3). The 18S was, therefore, the most suitable reference gene for real-time RT-PCR analysis of gene expression under all conditions. These results will support further studies on gene expression in P. neoaphidis. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-03-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822016000100259 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1016/j.bjm.2015.11.031 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
publisher.none.fl_str_mv |
Sociedade Brasileira de Microbiologia |
dc.source.none.fl_str_mv |
Brazilian Journal of Microbiology v.47 n.1 2016 reponame:Brazilian Journal of Microbiology instname:Sociedade Brasileira de Microbiologia (SBM) instacron:SBM |
instname_str |
Sociedade Brasileira de Microbiologia (SBM) |
instacron_str |
SBM |
institution |
SBM |
reponame_str |
Brazilian Journal of Microbiology |
collection |
Brazilian Journal of Microbiology |
repository.name.fl_str_mv |
Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM) |
repository.mail.fl_str_mv |
bjm@sbmicrobiologia.org.br||mbmartin@usp.br |
_version_ |
1752122208327565312 |