Identification and evaluation of reference genes for qRT-PCR normalization in ganoderma lucidum

Jiang Xu, Zhichao Xu, Yingjie Zhu, Hongmei Luo, Jun Qian, Aijia Ji, Yuanlei Hu, Wei Sun, Bo Wang, Jingyuan Song, Chao Sun, Shilin Chen

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Quantitative real-time reverse transcription PCR (qRT-PCR) is a rapid, sensitive, and reliable technique for gene expression studies. The accuracy and reliability of qRT-PCR results depend on the stability of the reference genes used for gene normalization. Therefore, a systematic process of reference gene evaluation is needed. Ganoderma lucidum is a famous medicinal mushroom in East Asia. In the current study, 10 potential reference genes were selected from the G. lucidum genomic data. The sequences of these genes were manually curated, and primers were designed following strict criteria. The experiment was conducted using qRT-PCR, and the stability of each candidate gene was assessed using four commonly used statistical programs - geNorm, NormFinder, BestKeeper, and RefFinder. According to our results, PP2A was expressed at the most stable levels under different fermentation conditions, and RPL4 was the most stably expressed gene in different tissues. RPL4, PP2A, and β-tubulin are the most commonly recommended reference genes for normalizing gene expression in the entire sample set. The current study provides a foundation for the further use of qRT-PCR in G. lucidum gene analysis.

Original languageEnglish (US)
Pages (from-to)120-126
Number of pages7
JournalCurrent Microbiology
Volume68
Issue number1
DOIs
StatePublished - Jan 1 2014
Externally publishedYes

ASJC Scopus subject areas

  • Microbiology
  • Applied Microbiology and Biotechnology

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    Xu, J., Xu, Z., Zhu, Y., Luo, H., Qian, J., Ji, A., Hu, Y., Sun, W., Wang, B., Song, J., Sun, C., & Chen, S. (2014). Identification and evaluation of reference genes for qRT-PCR normalization in ganoderma lucidum. Current Microbiology, 68(1), 120-126. https://doi.org/10.1007/s00284-013-0442-2