Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation
Identifieur interne : 001144 ( Main/Exploration ); précédent : 001143; suivant : 001145Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation
Auteurs : Jiying Wang ; Yanping Wang ; Huaizhong Wang ; Xiaojing Hao [République populaire de Chine] ; Ying Wu ; Jianfeng GuoSource :
- Asian-Australasian Journal of Animal Sciences [ 1011-2367 ] ; 2014.
Abstract
Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (
Url:
DOI: 10.5713/ajas.2013.13471
PubMed: 25049976
PubMed Central: 4093518
Affiliations:
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation</title><author><name sortKey="Wang, Jiying" sort="Wang, Jiying" uniqKey="Wang J" first="Jiying" last="Wang">Jiying Wang</name></author><author><name sortKey="Wang, Yanping" sort="Wang, Yanping" uniqKey="Wang Y" first="Yanping" last="Wang">Yanping Wang</name></author><author><name sortKey="Wang, Huaizhong" sort="Wang, Huaizhong" uniqKey="Wang H" first="Huaizhong" last="Wang">Huaizhong Wang</name></author><author><name sortKey="Hao, Xiaojing" sort="Hao, Xiaojing" uniqKey="Hao X" first="Xiaojing" last="Hao">Xiaojing Hao</name><affiliation wicri:level="1"><nlm:aff id="af1-ajas-27-4-471-3">Qingdao Institute of Animal Science and Veterinary Medicine, Qingdao 266100,<country>China</country>.</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wu, Ying" sort="Wu, Ying" uniqKey="Wu Y" first="Ying" last="Wu">Ying Wu</name></author><author><name sortKey="Guo, Jianfeng" sort="Guo, Jianfeng" uniqKey="Guo J" first="Jianfeng" last="Guo">Jianfeng Guo</name></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">25049976</idno><idno type="pmc">4093518</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4093518</idno><idno type="RBID">PMC:4093518</idno><idno type="doi">10.5713/ajas.2013.13471</idno><date when="2014">2014</date><idno type="wicri:Area/Pmc/Corpus">000812</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000812</idno><idno type="wicri:Area/Pmc/Curation">000812</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Curation">000812</idno><idno type="wicri:Area/Pmc/Checkpoint">000616</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Checkpoint">000616</idno><idno type="wicri:Area/Ncbi/Merge">000457</idno><idno type="wicri:Area/Ncbi/Curation">000457</idno><idno type="wicri:Area/Ncbi/Checkpoint">000457</idno><idno type="wicri:doubleKey">1011-2367:2014:Wang J:selection:of:reference</idno><idno type="wicri:Area/Main/Merge">001146</idno><idno type="wicri:Area/Main/Curation">001144</idno><idno type="wicri:Area/Main/Exploration">001144</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Selection of Reference Genes for Gene Expression Studies in Porcine Whole Blood and Peripheral Blood Mononuclear Cells under Polyinosinic:Polycytidylic Acid Stimulation</title><author><name sortKey="Wang, Jiying" sort="Wang, Jiying" uniqKey="Wang J" first="Jiying" last="Wang">Jiying Wang</name></author><author><name sortKey="Wang, Yanping" sort="Wang, Yanping" uniqKey="Wang Y" first="Yanping" last="Wang">Yanping Wang</name></author><author><name sortKey="Wang, Huaizhong" sort="Wang, Huaizhong" uniqKey="Wang H" first="Huaizhong" last="Wang">Huaizhong Wang</name></author><author><name sortKey="Hao, Xiaojing" sort="Hao, Xiaojing" uniqKey="Hao X" first="Xiaojing" last="Hao">Xiaojing Hao</name><affiliation wicri:level="1"><nlm:aff id="af1-ajas-27-4-471-3">Qingdao Institute of Animal Science and Veterinary Medicine, Qingdao 266100,<country>China</country>.</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea># see nlm:aff country strict</wicri:regionArea></affiliation></author><author><name sortKey="Wu, Ying" sort="Wu, Ying" uniqKey="Wu Y" first="Ying" last="Wu">Ying Wu</name></author><author><name sortKey="Guo, Jianfeng" sort="Guo, Jianfeng" uniqKey="Guo J" first="Jianfeng" last="Guo">Jianfeng Guo</name></author></analytic><series><title level="j">Asian-Australasian Journal of Animal Sciences</title><idno type="ISSN">1011-2367</idno><idno type="eISSN">1976-5517</idno><imprint><date when="2014">2014</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Investigating gene expression of immune cells of whole blood or peripheral blood mononuclear cells (PBMC) under polyinosinic:polycytidylic acid (poly I:C) stimulation is valuable for understanding the immune response of organism to RNA viruses. Quantitative real-time PCR (qRT-PCR) is a standard method for quantification of gene expression studies. However, the reliability of qRT-PCR data critically depends on proper selection of reference genes. In the study, using two different analysis programs, geNorm and NormFinder, we systematically evaluated the gene expression stability of six candidate reference genes (<italic>GAPDH</italic>, <italic>ACTB</italic>, <italic>B2M</italic>, <italic>RPL4</italic>, <italic>TBP</italic>, and <italic>PPIA</italic>) in samples of whole blood and PBMC with or without poly I:C stimulation. Generally, the six candidate genes performed a similar trend of expression stability in the samples of whole blood and PBMC, but more stably expressed in whole blood than in PBMC. geNorm ranked <italic>B2M</italic> and <italic>PPIA</italic> as the best combination for gene expression normalization, while according to NormFinder, <italic>TBP</italic> was ranked as the most stable reference gene, followed by <italic>B2M</italic> and <italic>PPIA</italic>. Comprehensively considering the results from the two programs, we recommended using the geometric mean of the three genes, <italic>TBP</italic>, <italic>PPIA</italic> and <italic>B2M</italic>, to normalize the gene expression of whole blood and PBMC with poly I:C stimulation. Our study is the first detailed survey of the gene expression stability in whole blood and PBMC with or without poly I:C stimulation and should be helpful for investigating the molecular mechanism involved in porcine whole blood and PBMC in response to poly I:C stimulation.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Andersen, Cl" uniqKey="Andersen C">CL Andersen</name></author><author><name sortKey="Jensen, Jl" uniqKey="Jensen J">JL Jensen</name></author><author><name sortKey=" Rntoft, Tf" uniqKey=" Rntoft T">TF Ørntoft</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Brym, P" uniqKey="Brym P">P Brym</name></author><author><name sortKey="Ru, A" uniqKey="Ru A">A Ruść</name></author><author><name sortKey="Kami Ski, S" uniqKey="Kami Ski S">S Kamiński</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cao, Y" uniqKey="Cao Y">Y Cao</name></author><author><name 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Wang</name><name sortKey="Wang, Yanping" sort="Wang, Yanping" uniqKey="Wang Y" first="Yanping" last="Wang">Yanping Wang</name><name sortKey="Wu, Ying" sort="Wu, Ying" uniqKey="Wu Y" first="Ying" last="Wu">Ying Wu</name></noCountry><country name="République populaire de Chine"><noRegion><name sortKey="Hao, Xiaojing" sort="Hao, Xiaojing" uniqKey="Hao X" first="Xiaojing" last="Hao">Xiaojing Hao</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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