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Evaluation of reference genes of Mytilus galloprovincialis and Ruditapes philippinarum infected with three bacteria strainsfor gene expression analysis

Published online by Cambridge University Press:  15 January 2015

Rebeca Moreira ,
Patricia Pereiro ,
María M. Costa ,
Antonio Figueras  and
Beatriz Novoa
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Abstract

Quantitative real-time polymerase chain reaction (qPCR) is probably the most used method for gene expression quantification because of its high sensitivity and specificity. Nevertheless, this technology can undergo experimental errors and variations. Normalization of the results using a reference gene is therefore necessary to minimize these variations. As the study of immune genes in bivalve mollusks has increased in the last years, the establishment of adequate and stable reference genes for bivalves is strongly required. We analyzed the behavior of four putative reference genes: ribosomal RNA 18S, actin, elongation factor 1 − α and α-tubulin. The suitability of these four genes as internal control for qPCR was evaluated in mussel (Mytilus galloprovincialis) and clam (Ruditapes philippinarum) hemocytes after bacterial challenge. Four independent approaches (BestKeeper, GeNorm, NormFinder and DeltaCt ) were used to assess the suitable genes for stable expression. For these particular circumstances, the most stable gene in hemocytes was elongation factor 1 − α for mussels and α-tubulin for clams.

Keywords

Shellfish immunityReference (housekeeping) genesqPCRMytilus galloprovincialisRuditapes philippinarumVibrio sppMicrococcus lysodeikticus
Type
Research Article
Information
Aquatic Living Resources , Volume 27 , Issue 3-4 , July 2014 , pp. 147 - 152
Copyright
© EDP Sciences, IFREMER, IRD 2015

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