English   español  
Please use this identifier to cite or link to this item: http://hdl.handle.net/10261/16218
Share/Impact:
Statistics
logo share SHARE logo core CORE   Add this article to your Mendeley library MendeleyBASE

Visualizar otros formatos: MARC | Dublin Core | RDF | ORE | MODS | METS | DIDL
Exportar a otros formatos:

Title

Evaluation of candidate reference genes for QPCR during ontogenesis and of immune-relevant tissues of European seabass (Dicentrarchus labrax)

AuthorsMitter, Karin; Kotoulas, Georgios; Magoulas, Antonios; Mulero, Victoriano; Sepulcre, Pilar; Figueras Huerta, Antonio ; Novoa, Beatriz ; Sarropoulou, Elena
KeywordsqRT-PCR
Reference gene
Aquaculture
Dicentrarchus labrax
Developmental stage
Infection
Nodavirus
Vibrio anguillarum
Issue Date4-May-2009
PublisherElsevier
CitationComparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 153(4): 340-347 (2009)
AbstractThe expression level of mRNA can vary significantly in different experimental conditions, such as stress, infection, developmental stage or tissue. Suitable reference genes are expected to exhibit constant expression levels. However no single gene is constitutively expressed in all cell types and under all experimental conditions. It has become clear that expression stability of the intended reference gene has to be examined before each experiment. For expression studies using quantitative real-time PCR (qPCR) at least two reference genes have to be applied. So far expression studies in the European seabass (Dicentrarchus labrax) as well as in the Gilthead seabream (Sparus aurata) have been performed with only one reference gene (S18, Ef-1 alpha or Gapdh). Though significant variations showed up in other teleost species such as the Atlantic halibut and the zebrafish affirming the need for proper normalization strategies, the present study aims at identifying suitable reference genes among nine candidates [glyceraldehyde-phosphate-dehydrogenase (Gapdh), β-actin (two regions of β-actin), 40S ribosomal protein S30 (Fau), ribosomal protein L13 a (L13a), β2-tubulin (Tubb2) and tyrosine 3 monooxygenase/tryptophan 5-monooxygenase activation protein (Tyr)] for expression analysis of 8 developmental stages and a tissue panel (spleen, liver, kidney and brain) with samples infected with Nodavirus and Vibrio anguillarum in D. labrax. Besides the analysis of raw Ct-values, the gene expression stability was determined using two different software applications BestKeeper and NormFinder. According to both algorithms the best two reference genes for an appropriate normalization approach during D. labrax development are Ef-1 alpha and L13a whereas in the tissue panel Fau and L13a are recommended for qPCR normalization.
Description8 pages, 6 figures, 5 tables.-- PMID: 19398033 [PubMed].-- Printed version published Aug 2009.
Corrigendum published in Comp Biochem Physiol B Biochem Mol Biol. Aug 6, 2009, http://dx.doi.org/10.1016/j.cbpb.2009.07.008
Publisher version (URL)http://dx.doi.org/10.1016/j.cbpb.2009.04.009
URIhttp://hdl.handle.net/10261/16218
DOI10.1016/j.cbpb.2009.04.009
ISSN1096-4959
Appears in Collections:(IIM) Artículos
Files in This Item:
There are no files associated with this item.
Show full item record
Review this work
 

Related articles:


WARNING: Items in Digital.CSIC are protected by copyright, with all rights reserved, unless otherwise indicated.