Evaluation of reference genes for gene expression studies using quantitative real-time PCR in Drosophila melanogaster after chemical exposures

Yeong Ho Kim, Yi Seul Kim, Young Ho Kim

Research output: Contribution to journalArticlepeer-review

11 Scopus citations

Abstract

The fruit fly, Drosophila melanogaster, is mainly found in fermented and rotten fruits and is more tolerant to chemicals emitted during the fermentation process than other organisms. Its distinctive habitat suggests an evolutionary adaptation to the chemicals, such as acetic acid, ethanol, and 2-phenylethanol. Quantitative real-time PCR (qRT-PCR) assays can be performed to analyze the expression patterns of D. melanogaster genes putatively involved in chemical detoxification and metabolism, to better understand D. melanogaster adaptions to its environment. The selection of stably-expressed, internal reference genes across samples is crucial for accurate normalization of target gene expression parameters. In this study, we investigated the transcription levels of ten candidate reference genes: hsp22, ef1β, α-tub, rpL18, rpS3, argk, nd, tbp, gapdh, and ace, in D. melanogaster exposed to various concentrations of acetic acid, ethanol, and 2-phenylethanol, and expression stability of these genes was evaluated using three different programs: geNorm, NormFinder, and BestKeeper. These three software resulted in different stable genes, but suggested the selection of multiple reference genes for target gene normalization. For validation of reference genes, expression levels of adh were normalized with different references and combination of multiple genes. In the flies exposed to three chemicals, rpL18 was commonly suggested to be used for the most stable reference gene for comparing expression levels of genes potentially associated with environmental chemical tolerance in D. melanogaster.

Original languageEnglish
Pages (from-to)385-394
Number of pages10
JournalJournal of Asia-Pacific Entomology
Volume23
Issue number2
DOIs
StatePublished - Jun 2020

Keywords

  • Acetic acid
  • Drosophila melanogaster
  • Ethanol, 2-phenylethanol
  • Quantitative real-time PCR
  • Reference gene

Fingerprint

Dive into the research topics of 'Evaluation of reference genes for gene expression studies using quantitative real-time PCR in Drosophila melanogaster after chemical exposures'. Together they form a unique fingerprint.

Cite this