Selection of reference genes for quantitative real-time PCR analysis in chicken embryo fibroblasts infected with avian leukosis virus subgroup J
Identifieur interne : 001225 ( Main/Exploration ); précédent : 001224; suivant : 001226Selection of reference genes for quantitative real-time PCR analysis in chicken embryo fibroblasts infected with avian leukosis virus subgroup J
Auteurs : Falong Yang [République populaire de Chine] ; Xiaowen Lei [République populaire de Chine] ; Alexander Rodriguez-Palacios [États-Unis] ; Cheng Tang [République populaire de Chine] ; Hua Yue [République populaire de Chine]Source :
- BMC Research Notes [ 1756-0500 ] ; 2013.
Abstract
The selection of stably expressed reference genes is a prerequisite when evaluating gene expression, via real-time PCR, in cells in response to viral infections. The objective of our study was to identify suitable reference genes for mRNA expression analysis in chicken embryonic fibroblasts (CEF) after infection with avian leukosis virus subgroup J (ALV-J).
The expression levels of 11 potential reference genes in CEF infected with ALV-J were determined by real-time PCR. The expression stability of these genes were analyzed and ranked using the geNorm tool. Analysis indicated that the genes
The
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DOI: 10.1186/1756-0500-6-402
PubMed: 24099561
PubMed Central: 3851545
Affiliations:
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Nationalities, Chengdu 610041</wicri:regionArea><wicri:noRegion>Chengdu 610041</wicri:noRegion></affiliation></author><author><name sortKey="Lei, Xiaowen" sort="Lei, Xiaowen" uniqKey="Lei X" first="Xiaowen" last="Lei">Xiaowen Lei</name><affiliation wicri:level="1"><nlm:aff id="I2">Ganzhou Institute of Animal Science, Ganzhou 341401, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Ganzhou Institute of Animal Science, Ganzhou 341401</wicri:regionArea><wicri:noRegion>Ganzhou 341401</wicri:noRegion></affiliation></author><author><name sortKey="Rodriguez Palacios, Alexander" sort="Rodriguez Palacios, Alexander" uniqKey="Rodriguez Palacios A" first="Alexander" last="Rodriguez-Palacios">Alexander Rodriguez-Palacios</name><affiliation wicri:level="2"><nlm:aff id="I3">Food Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH 44691, USA</nlm:aff><country xml:lang="fr">États-Unis</country><wicri:regionArea>Food Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH 44691</wicri:regionArea><placeName><region type="state">Ohio</region></placeName></affiliation></author><author><name sortKey="Tang, Cheng" sort="Tang, Cheng" uniqKey="Tang C" first="Cheng" last="Tang">Cheng Tang</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041</wicri:regionArea><wicri:noRegion>Chengdu 610041</wicri:noRegion></affiliation></author><author><name sortKey="Yue, Hua" sort="Yue, Hua" uniqKey="Yue H" first="Hua" last="Yue">Hua Yue</name><affiliation wicri:level="1"><nlm:aff id="I1">Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Veterinary Medicine, College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041</wicri:regionArea><wicri:noRegion>Chengdu 610041</wicri:noRegion></affiliation></author></analytic><series><title level="j">BMC Research Notes</title><idno type="eISSN">1756-0500</idno><imprint><date when="2013">2013</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>Background</title><p>The selection of stably expressed reference genes is a prerequisite when evaluating gene expression, via real-time PCR, in cells in response to viral infections. The objective of our study was to identify suitable reference genes for mRNA expression analysis in chicken embryonic fibroblasts (CEF) after infection with avian leukosis virus subgroup J (ALV-J).</p></sec><sec><title>Findings</title><p>The expression levels of 11 potential reference genes in CEF infected with ALV-J were determined by real-time PCR. The expression stability of these genes were analyzed and ranked using the geNorm tool. Analysis indicated that the genes <italic>RPL30</italic> (ribosomal protein L30) and <italic>SDHA</italic> (succinate dehydrogenase complex, subunit A) were the most stably expressed genes in the ALV-J infected CEF.</p></sec><sec><title>Conclusions</title><p>The <italic>RPL30</italic> and <italic>SDHA</italic> were deemed suitable for use as reference genes for real-time PCR analysis of mRNA gene expression during ALV-J infection, whereas commonly used <italic>ACTB</italic> and <italic>GAPDH</italic> are unsuitable to be reference genes.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Wang, F" uniqKey="Wang F">F Wang</name></author><author><name sortKey="Wang, X" uniqKey="Wang X">X Wang</name></author><author><name sortKey="Chen, X" uniqKey="Chen X">X Chen</name></author><author><name sortKey="Liu, X" uniqKey="Liu X">X Liu</name></author><author><name sortKey="Cheng, Z" uniqKey="Cheng Z">Z 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uniqKey="Van Roy N">N Van Roy</name></author><author><name sortKey="De Paepe, A" uniqKey="De Paepe A">A De Paepe</name></author><author><name sortKey="Speleman, F" uniqKey="Speleman F">F Speleman</name></author></analytic></biblStruct></listBibl></div1></back></TEI><affiliations><list><country><li>République populaire de Chine</li><li>États-Unis</li></country><region><li>Ohio</li></region></list><tree><country name="République populaire de Chine"><noRegion><name sortKey="Yang, Falong" sort="Yang, Falong" uniqKey="Yang F" first="Falong" last="Yang">Falong Yang</name></noRegion><name sortKey="Lei, Xiaowen" sort="Lei, Xiaowen" uniqKey="Lei X" first="Xiaowen" last="Lei">Xiaowen Lei</name><name sortKey="Tang, Cheng" sort="Tang, Cheng" uniqKey="Tang C" first="Cheng" last="Tang">Cheng Tang</name><name sortKey="Yue, Hua" sort="Yue, Hua" uniqKey="Yue H" first="Hua" last="Yue">Hua Yue</name></country><country name="États-Unis"><region name="Ohio"><name sortKey="Rodriguez Palacios, Alexander" 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