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A Panel of Stably E...
A Panel of Stably Expressed Reference Genes for Real-Time qPCR Gene Expression Studies of Mallards (Anas platyrhynchos)
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- Chapman, Joanne R. (författare)
- Linnéuniversitetet,Institutionen för biologi och miljö (BOM),Ctr Ecol & Evolut Microbial Model Syst EEMiS
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- Helin, Anu S. (författare)
- Linnéuniversitetet,Institutionen för biologi och miljö (BOM),Ctr Ecol & Evolut Microbial Model Syst EEMiS
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- Wille, Michelle (författare)
- Uppsala universitet,Linnéuniversitetet,Institutionen för biologi och miljö (BOM),Uppsala University,Ctr Ecol & Evolut Microbial Model Syst EEMiS,Institutionen för medicinsk biokemi och mikrobiologi,Centre for Ecology and Evolution in Microbial Model Systems, Linnaeus University
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- Atterby, Clara (författare)
- Uppsala universitet,Institutionen för medicinsk biokemi och mikrobiologi
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- Järhult, Josef D., 1975- (författare)
- Uppsala universitet,Infektionssjukdomar,Institutionen för medicinsk biokemi och mikrobiologi
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- Fridlund, Jimmy (författare)
- Linnéuniversitetet,Institutionen för biologi och miljö (BOM),Ctr Ecol & Evolut Microbial Model Syst EEMiS
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- Waldenström, Jonas (författare)
- Linnéuniversitetet,Institutionen för biologi och miljö (BOM),Ctr Ecol & Evolut Microbial Model Syst EEMiS
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(creator_code:org_t)
- 2016-02-17
- 2016
- Engelska.
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Ingår i: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 11:2
- Relaterad länk:
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https://doi.org/10.1...
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https://uu.diva-port... (primary) (Raw object)
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- Determining which reference genes have the highest stability, and are therefore appropriate for normalising data, is a crucial step in the design of real-time quantitative PCR (qPCR) gene expression studies. This is particularly warranted in non-model and ecologically important species for which appropriate reference genes are lacking, such as the mallard-a key reservoir of many diseases with relevance for human and livestock health. Previous studies assessing gene expression changes as a consequence of infection in mallards have nearly universally used beta-actin and/or GAPDH as reference genes without confirming their suitability as normalisers. The use of reference genes at random, without regard for stability of expression across treatment groups, can result in erroneous interpretation of data. Here, eleven putative reference genes for use in gene expression studies of the mallard were evaluated, across six different tissues, using a low pathogenic avian influenza A virus infection model. Tissue type influenced the selection of reference genes, whereby different genes were stable in blood, spleen, lung, gastrointestinal tract and colon. beta-actin and GAPDH generally displayed low stability and are therefore inappropriate reference genes in many cases. The use of different algorithms (GeNorm and NormFinder) affected stability rankings, but for both algorithms it was possible to find a combination of two stable reference genes with which to normalise qPCR data in mallards. These results highlight the importance of validating the choice of normalising reference genes before conducting gene expression studies in ducks. The fact that nearly all previous studies of the influence of pathogen infection on mallard gene expression have used a single, non-validated reference gene is problematic. The toolkit of putative reference genes provided here offers a solid foundation for future studies of gene expression in mallards and other waterfowl.
Ämnesord
- NATURVETENSKAP -- Biologi -- Ekologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Ecology (hsv//eng)
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Infektionsmedicin (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Infectious Medicine (hsv//eng)
Nyckelord
- Zoonotic Ecology
- Zoonotisk ekologi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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