Evaluation of putative internal reference genes for gene expression normalization in Nannochloropsis sp by quantitative real-time RT-PCR

文献类型： 外文期刊

第一作者： Cao, Shaona

作者： Cao, Shaona;Zhang, Xiaowen;Ye, Naihao;Fan, Xiao;Xu, Dong;Cao, Shaona;Wang, Yitao;Wang, Wenqi;Liang, Chengwei

作者机构：

关键词： Nannochloropsis sp.;Gene expression;Normalization;RT-qPCR;Reference genes

期刊名称：BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS （ 影响因子：3.575； 五年影响因子：3.381 ）

ISSN： 0006-291X

年卷期： 2012 年 424 卷 1 期

页码：

收录情况： SCI

摘要： Quantitative real-time reverse transcription PCR (RT-qPCR), a sensitive technique for quantifying gene expression, depends on the stability of the reference gene(s) used for data normalization. To date, few studies on reference genes have been undertaken for Nannochloropsis sp. In this study, 12 potential reference genes were evaluated for their expression stability using the geNorm and NormFinder statistical algorithms by RT-qPCR. The results showed that the best reference genes differed depending on the treatments: different light intensities (DL), the diurnal cycle (DC), high light intensity (HL) and low temperature treatments (LT). A combination of ACT1, ACT2 and TUA would be appropriate as a reference panel for normalizing gene expression data across all the treatments. ACT2 showed the most stable expression across all tested samples but was not the most stable one for individual treatments. Though 18S showed the least stable expression considering all tested samples, it is the most stable one for LT using geNorm. The expression of Lhc confirmed that the appropriate reference genes are crucial. These results provide a foundation for more accurate use of RT-qPCR under different experimental conditions in Nannochloropsis sp. gene analysis. (C) 2012 Elsevier Inc. All rights reserved.

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