Evaluation of potential reference genes for reverse transcription-qPCR studies of physiological responses in Drosophila melanogaster

Ponton Fleur, Chapuis Marie Pierre, Pernice Mathieu, Sword Gregory A., Simpson Stephen J.. 2011. Evaluation of potential reference genes for reverse transcription-qPCR studies of physiological responses in Drosophila melanogaster. Journal of Insect Physiology, 57 (6) : pp. 840-850.

Quartile : Q1, Sujet : ENTOMOLOGY / Quartile : Q2, Sujet : PHYSIOLOGY

Liste HCERES des revues (en SHS) : oui

Thème(s) HCERES des revues (en SHS) : Psychologie-éthologie-ergonomie

Abstract : Drosophila melanogaster is one of the most important genetic models and techniques such as reverse transcription quantitative real-time PCR (RT-qPCR) are being employed extensively for deciphering the genetics basis of physiological functions. In RT-qPCR, the expression levels of target genes are estimated on the basis of endogenous controls. The purpose of these reference genes is to control for variations in RNA quantity and quality. Although determination of suitable reference genes is essential to RT-qPCR studies, reports on the evaluation of reference genes in D. melanogaster studies are lacking. We analyzed the expression levels of seven candidate reference genes (Actin, EF1, Mnf, Rps20, Rpl32, Tubulin and 18S) in flies that were injured, heat-stressed, or fed different diets. Statistical analyses of variation were determined using three established software programs for reference gene selection, geNorm, NormFinder and BestKeeper. Best-ranked references genes differed across the treatments. Normalization candidacy of the selected candidate reference genes was supported by an analysis of gene expression values obtained from microarray datasets available online. The differences between the experimental treatments suggest that assessing the stability of reference gene expression patterns, determining candidates and testing their suitability is required for each experimental investigation. (Résumé d'auteur)

Mots-clés Agrovoc : Drosophila melanogaster, Transcription inverse, Stress thermique, Gène

Classification Agris : L10 - Animal genetics and breeding
L50 - Animal physiology and biochemistry

Champ stratégique Cirad : Hors axes (2005-2013)

Auteurs et affiliations

• Ponton Fleur, University of Sydney (AUS)
• Chapuis Marie Pierre, CIRAD-BIOS-UPR Bioagresseurs (FRA)
• Pernice Mathieu, University of Queensland (AUS)
• Sword Gregory A., University of Sydney (AUS)
• Simpson Stephen J., University of Sydney (AUS)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/561939/)

Metrics

[ Page générée et mise en cache le 2021-03-02 ]