Exploration of Reference Genes for Quantitative Real-Time PCR in marine diatom Phaeodactylum tricornutum
- Exploration of Reference Genes for Quantitative Real-Time PCR in marine diatom Phaeodactylum tricornutum
- 정유진; 신복규; 판철호; 김상민
- Phaeodactylum tricornutum; housekeeping genes; qRT-PCR; diatom
- Issue Date
- 2015년도 한국응용생명화학회 국제학술대회
- Phaeodactylum tricornutum is a diatom whose genome information is available. However, research of target gene expression in this microalgae is not easy due to the absence of confident reference genes. In this study, we investigated the expression levels of five potential reference genes, Actin16, RPS (ribosomal protein small subunit), Cdk4 (Cyclin-dependent kinase 4), Histone H4, and TBP (TATA box binding protein) in P. tricornutum cells by quantitative real-time PCR(qRT-PCR) in order to select proper genes as internal standards for gene expression. P. tricornutum was grown under several different conditions and RNA samples were isolated from P. tricornutum cells harvested under different growth stages, culture conditions. Among the tested genes, the expression profiles of Histone H4 showed the best stability during growth phase and light/dark conditions, suggesting to be the most suitable candidate for housekeeping genes in P. tricornutum. As positive control experiment, we further checked RBC (RuBisCo) gene expression in P. tricorntum cells grown under silicon starvation condition by using Histone H4 as reference gene and confirmed the same result as reported data. These results suggest that Histone H4 and Cdk4 were the most reliable reference gene for qRT-PCR in themarine diatom P. tricornutum.
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