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Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis
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 can...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822748/ https://www.ncbi.nlm.nih.gov/pubmed/26887253 http://dx.doi.org/10.1016/j.bjm.2015.11.031 |
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author | Chen, Chun Xie, Tingna Ye, Sudan Jensen, Annette Bruun Eilenberg, Jørgen |
author_facet | Chen, Chun Xie, Tingna Ye, Sudan Jensen, Annette Bruun Eilenberg, Jørgen |
author_sort | Chen, Chun |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4822748 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-48227482016-05-17 Selection of reference genes for expression analysis in the entomophthoralean fungus Pandora neoaphidis Chen, Chun Xie, Tingna Ye, Sudan Jensen, Annette Bruun Eilenberg, Jørgen Braz J Microbiol Genetics and Molecular Microbiology 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. Elsevier 2016-02-17 /pmc/articles/PMC4822748/ /pubmed/26887253 http://dx.doi.org/10.1016/j.bjm.2015.11.031 Text en © 2015 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Genetics and Molecular Microbiology Chen, Chun Xie, Tingna Ye, Sudan Jensen, Annette Bruun Eilenberg, Jørgen 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 |
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_short | 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 |
topic | Genetics and Molecular Microbiology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4822748/ https://www.ncbi.nlm.nih.gov/pubmed/26887253 http://dx.doi.org/10.1016/j.bjm.2015.11.031 |
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