Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon
Identifieur interne : 001001 ( Main/Exploration ); précédent : 001000; suivant : 001002Evaluation of potential reference genes in real-time RT-PCR studies of Atlantic salmon
Auteurs : P L A. Olsvik [Norvège] ; Kai K. Lie [Norvège] ; Ann-Elise O. Jordal [Norvège] ; Tom O. Nilsen [Norvège] ; Ivar Hordvik [Norvège]Source :
- BMC Molecular Biology [ 1471-2199 ] ; 2005.
Abstract
Salmonid fishes are among the most widely studied model fish species but reports on systematic evaluation of reference genes in qRT-PCR studies is lacking.
The stability of six potential reference genes was examined in eight tissues of Atlantic salmon (
Overall, this work suggests that the EF1AA and EF1AB genes can be useful as reference genes in qRT-PCR examination of gene expression in the Atlantic salmon.
Url:
DOI: 10.1186/1471-2199-6-21
PubMed: 16293192
PubMed Central: 1314898
Affiliations:
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Le document en format XML
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<front><div type="abstract" xml:lang="en"><sec><title>Background</title>
<p>Salmonid fishes are among the most widely studied model fish species but reports on systematic evaluation of reference genes in qRT-PCR studies is lacking.</p>
</sec>
<sec><title>Results</title>
<p>The stability of six potential reference genes was examined in eight tissues of Atlantic salmon (<italic>Salmo salar</italic>
), to determine the most suitable genes to be used in quantitative real-time RT-PCR analyses. The relative transcription levels of genes encoding 18S rRNA, S20 ribosomal protein, β-actin, glyceraldehyde-3P-dehydrogenase (GAPDH), and two paralog genes encoding elongation factor 1A (EF1A<sub>A </sub>
and EF1A<sub>B</sub>
) were quantified in gills, liver, head kidney, spleen, thymus, brain, muscle, and posterior intestine in six untreated adult fish, in addition to a group of individuals that went through smoltification. Based on calculations performed with the <italic>geNorm </italic>
VBA applet, which determines the most stable genes from a set of tested genes in a given cDNA sample, the ranking of the examined genes in adult Atlantic salmon was EF1A<sub>B</sub>
>EF1A<sub>A</sub>
>β-actin>18S rRNA>S20>GAPDH. When the same calculations were done on a total of 24 individuals from four stages in the smoltification process (presmolt, smolt, smoltified seawater and desmoltified freshwater), the gene ranking was EF1A<sub>B</sub>
>EF1A<sub>A</sub>
>S20>β-actin>18S rRNA>GAPDH.</p>
</sec>
<sec><title>Conclusion</title>
<p>Overall, this work suggests that the EF1A<sub>A </sub>
and EF1A<sub>B </sub>
genes can be useful as reference genes in qRT-PCR examination of gene expression in the Atlantic salmon.</p>
</sec>
</div>
</front>
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