Department of Green Life Science, Sangmyung University, Seoul 110-743, South Korea.
Eur J Protistol. 2012 Aug;48(3):199-206. doi: 10.1016/j.ejop.2011.11.001. Epub 2011 Dec 29.
Housekeeping genes (HKGs) are required for the normalization of expression levels in real-time PCR, and their selection is critical for gene expression studies. However, stable expressions of candidate HKGs vary among organisms and tissues and even according to environmental conditions. Here, we evaluated the gene expressions of 10 frequently used HKGs, including 18S rRNA, P2, ACT, TUA, TUB, CYC, eIF4E, MDH, UBQ, and GAPDH, with a total of 17 RNA samples extracted from the dinoflagellate Prorocentrum minimum. All the RNAs were prepared from various cells cultured under different conditions, including thermal shocks, toxic chemical exposures, and different life stages. Via C(T) analyses of the 10 HKGs using the geNorm software, TUA was selected as the most stable HKG for gene expression studies of the dinoflagellate, followed by MDH. Pair-wise variation (V) analysis showed that at least 2 genes were required for accurate normalization of gene expression studies depending on the experimental conditions. These HKGs are useful internal controls for the normalization of gene expression in P. minimum.
管家基因（HKGs）是实时 PCR 中表达水平标准化所必需的，其选择对于基因表达研究至关重要。然而，候选 HKGs 的稳定表达在不同的生物体和组织中甚至根据环境条件而有所不同。在这里，我们评估了 10 种常用 HKGs 的基因表达，包括 18S rRNA、P2、ACT、TUA、TUB、CYC、eIF4E、MDH、UBQ 和 GAPDH，总共从原甲藻 Prorocentrum minimum 中提取了 17 个 RNA 样本。所有 RNA 均来自不同细胞培养物，包括热冲击、有毒化学物质暴露和不同的生命阶段。通过 geNorm 软件对 10 个 HKGs 的 C(T)分析，TUA 被选为原甲藻基因表达研究最稳定的 HKG，其次是 MDH。成对差异（V）分析表明，根据实验条件，至少需要 2 个基因才能准确归一化基因表达研究。这些 HKGs 是 P. minimum 中基因表达标准化的有用内参。
