An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either Piscirickettsia salmonis or IPNV
Identifieur interne : 001480 ( Main/Exploration ); précédent : 001479; suivant : 001481An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either Piscirickettsia salmonis or IPNV
Auteurs : Andrea A. Pe A [Chili] ; Niels C. Bols [Canada] ; Sergio H. Marshall [Chili]Source :
- BMC Research Notes [ 1756-0500 ] ; 2010.
Abstract
Due to the limited number of species specific antibodies against fish proteins, differential gene expression analyses are vital for the study of host immune responses. Quantitative real-time reverse transcription PCR (qRT-PCR) is one of the most powerful tools for this purpose. Nevertheless, the accuracy of the method will depend on the careful selection of genes whose expression are stable and can be used as internal controls for a particular experimental setting.
The expression stability of five commonly used housekeeping genes [beta-actin (
Based on the data presented here with the cell culture models CHSE-214 and RTS11, we suggest the initial choice of
Url:
DOI: 10.1186/1756-0500-3-101
PubMed: 20398263
PubMed Central: 2873344
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">An evaluation of potential reference genes for stability of expression in two salmonid cell lines after infection with either <italic>Piscirickettsia salmonis </italic>or IPNV</title><author><name sortKey="Pe A, Andrea A" sort="Pe A, Andrea A" uniqKey="Pe A A" first="Andrea A" last="Pe A">Andrea A. Pe A</name><affiliation wicri:level="1"><nlm:aff id="I1">Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso, Chile</nlm:aff><country xml:lang="fr">Chili</country><wicri:regionArea>Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso</wicri:regionArea><wicri:noRegion>Valparaíso</wicri:noRegion></affiliation></author><author><name sortKey="Bols, Niels C" sort="Bols, Niels C" uniqKey="Bols N" first="Niels C" last="Bols">Niels C. Bols</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1</wicri:regionArea><wicri:noRegion>ON N2L 3G1</wicri:noRegion></affiliation></author><author><name sortKey="Marshall, Sergio H" sort="Marshall, Sergio H" uniqKey="Marshall S" first="Sergio H" last="Marshall">Sergio H. Marshall</name><affiliation wicri:level="1"><nlm:aff id="I1">Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso, Chile</nlm:aff><country xml:lang="fr">Chili</country><wicri:regionArea>Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. 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Pe A</name><affiliation wicri:level="1"><nlm:aff id="I1">Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso, Chile</nlm:aff><country xml:lang="fr">Chili</country><wicri:regionArea>Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso</wicri:regionArea><wicri:noRegion>Valparaíso</wicri:noRegion></affiliation></author><author><name sortKey="Bols, Niels C" sort="Bols, Niels C" uniqKey="Bols N" first="Niels C" last="Bols">Niels C. Bols</name><affiliation wicri:level="1"><nlm:aff id="I2">Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, Canada</nlm:aff><country xml:lang="fr">Canada</country><wicri:regionArea>Department of Biology, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1</wicri:regionArea><wicri:noRegion>ON N2L 3G1</wicri:noRegion></affiliation></author><author><name sortKey="Marshall, Sergio H" sort="Marshall, Sergio H" uniqKey="Marshall S" first="Sergio H" last="Marshall">Sergio H. Marshall</name><affiliation wicri:level="1"><nlm:aff id="I1">Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso, Chile</nlm:aff><country xml:lang="fr">Chili</country><wicri:regionArea>Laboratorio de Genética e Inmunología Molecular, Instituto de Biología, Pontificia Universidad Católica de Valparaíso, Av. Brasil 2950, Valparaíso</wicri:regionArea><wicri:noRegion>Valparaíso</wicri:noRegion></affiliation></author></analytic><series><title level="j">BMC Research Notes</title><idno type="eISSN">1756-0500</idno><imprint><date when="2010">2010</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>Due to the limited number of species specific antibodies against fish proteins, differential gene expression analyses are vital for the study of host immune responses. Quantitative real-time reverse transcription PCR (qRT-PCR) is one of the most powerful tools for this purpose. Nevertheless, the accuracy of the method will depend on the careful selection of genes whose expression are stable and can be used as internal controls for a particular experimental setting.</p></sec><sec><title>Findings</title><p>The expression stability of five commonly used housekeeping genes [beta-actin (<italic>ACTB</italic>), elongation factor 1-alpha (<italic>EF1A</italic>), ubiquitin (<italic>UBQ</italic>), glyceraldehyd-3-phosphate dehydrogenase (<italic>GAPDH</italic>) and tubulin alpha (<italic>TUBA</italic>)] were monitored in salmonid cell lines CHSE-214 and RTS11 after infection with two of the most fastidious fish pathogens, the facultative bacterium <italic>Piscirickettsia salmonis </italic>and the aquabirnavirus IPNV (Infectious Pancreatic Necrosis Virus). After geNorm analysis, <italic>UBQ </italic>and <italic>EF1A </italic>appeared as the most stable, although <italic>EF1A </italic>was slightly upregulated at late stages of <italic>P. salmonis </italic>infection in RTS11. <italic>ACTB </italic>instead, showed a good performance in each case, being always considered within the three most stable genes of the panel. In contrast, infection-dependent differential regulation of <italic>GAPDH </italic>and <italic>TUBA </italic>was also demonstrated.</p></sec><sec><title>Conclusion</title><p>Based on the data presented here with the cell culture models CHSE-214 and RTS11, we suggest the initial choice of <italic>UBQ</italic>, <italic>ACTB </italic>and <italic>EF1A </italic>as reference genes in qRT-PCR assays for studying the effect of <italic>P. salmonis </italic>and IPNV on the host immune response.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Bustin, Sa" uniqKey="Bustin S">SA Bustin</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Huggett, J" uniqKey="Huggett J">J Huggett</name></author><author><name sortKey="Dheda, K" uniqKey="Dheda K">K Dheda</name></author><author><name sortKey="Bustin, S" uniqKey="Bustin S">S Bustin</name></author><author><name sortKey="Zumla, A" uniqKey="Zumla A">A 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Pe A</name></noRegion><name sortKey="Marshall, Sergio H" sort="Marshall, Sergio H" uniqKey="Marshall S" first="Sergio H" last="Marshall">Sergio H. Marshall</name></country><country name="Canada"><noRegion><name sortKey="Bols, Niels C" sort="Bols, Niels C" uniqKey="Bols N" first="Niels C" last="Bols">Niels C. Bols</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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