Selection and validation of reference genes for qRT-PCR expression analysis of candidate genes involved in olfactory communication in the butterfly Bicyclus anynana

PloS One
Alok ArunCaroline M Nieberding

Abstract

Real-time quantitative reverse transcription PCR (qRT-PCR) is a technique widely used to quantify the transcriptional expression level of candidate genes. qRT-PCR requires the selection of one or several suitable reference genes, whose expression profiles remain stable across conditions, to normalize the qRT-PCR expression profiles of candidate genes. Although several butterfly species (Lepidoptera) have become important models in molecular evolutionary ecology, so far no study aimed at identifying reference genes for accurate data normalization for any butterfly is available. The African bush brown butterfly Bicyclus anynana has drawn considerable attention owing to its suitability as a model for evolutionary ecology, and we here provide a maiden extensive study to identify suitable reference gene in this species. We monitored the expression profile of twelve reference genes: eEF-1α, FK506, UBQL40, RpS8, RpS18, HSP, GAPDH, VATPase, ACT3, TBP, eIF2 and G6PD. We tested the stability of their expression profiles in three different tissues (wings, brains, antennae), two developmental stages (pupal and adult) and two sexes (male and female), all of which were subjected to two food treatments (food stress and control feeding ad libi...Continue Reading

References

Jul 1, 1997·Mammalian Genome : Official Journal of the International Mammalian Genome Society·M S LyuC A Kozak
Oct 26, 2001·Current Biology : CB·C R BrunettiS B Carroll
Jun 4, 2002·Nature Reviews. Genetics·Patrícia Beldade, Paul M Brakefield
Jan 7, 2004·Biochemical and Biophysical Research Communications·Aleksandar RadonićAndreas Nitsche
Apr 9, 2005·Genes and Immunity·J HuggettA Zumla
Jul 29, 2005·Proceedings. Biological Sciences·Kendra A Robertson, Antónia Monteiro
Feb 10, 2007·Heredity·P BeldadeA Papanicolaou
Jul 24, 2008·PloS One·Caroline M NieberdingPaul M Brakefield
Sep 25, 2008·The American Naturalist·P M BrakefieldP B Koch
Jun 16, 2009·Insect Molecular Biology·D GonzálezR Renthal
Dec 2, 2010·Experimental Gerontology·Jeroen PijpeBas J Zwaan
Dec 22, 2010·Molecular Biology and Evolution·Lino OmettoLaurent Keller
Mar 7, 2012·Ecology Letters·Caroline M NieberdingPaul M Brakefield
Jun 23, 2012·Nature·UNKNOWN Heliconius Genome Consortium
Aug 22, 2012·Nature Reviews. Gastroenterology & Hepatology·Claire Greenhill
Mar 8, 2013·Proceedings. Biological Sciences·Erik van BergenCaroline M Nieberding
Feb 7, 2014·Proceedings. Biological Sciences·Astrid T GrootDavid G Heckel
May 28, 2014·Nature Communications·Marjorie A LiénardChrister Löfstedt
Mar 6, 2015·Proceedings. Biological Sciences·P M B BacquetC M Nieberding

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Methods Mentioned

BETA
reverse transcription PCR
PCR
electrophoresis

Software Mentioned

geNorm
R
NormFinder
BoxPlotR
Primer
Gene Runner
TBLASTN

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