| 实时荧光定量PCR分析中不同铁浓度处理下玛氏骨条藻内参基因的筛选 |
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| 引用本文： | 张梅,邢永泽,甄毓,米铁柱,于志刚. 实时荧光定量PCR分析中不同铁浓度处理下玛氏骨条藻内参基因的筛选[J]. 海洋学报, 2020, 42(2): 124-133. DOI: 10.3969/j.issn.0253-4193.2020.02.013 |
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| 作者姓名： | 张梅 邢永泽 甄毓 米铁柱 于志刚 |
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| 作者单位： | 1.中国海洋大学 海洋环境与生态教育部重点实验室，山东 青岛 266100 |
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| 基金项目： | 国家重点研发计划项目（2017YFC1404402）；鳌山科技创新计划项目（2016ASKJ02）。 |
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| 摘 要： | 实时荧光定量PCR(qRT-PCR)是定量分析基因表达的常用方法,选择合适的内参基因对准确分析目的基因表达水平至关重要。本研究以不同铁浓度培养条件下的玛氏骨条藻为材料,定量分析Cytb、EF-1α、HPRT、UBC、GAPDH、β-actin以及β-tubulin 7个内参基因的表达情况,并利用GeNorm、NormFinder和BestKeeper软件对这些内参基因的稳定性进行综合评价。结果表明,Cytb和EF-1α的表达稳定性较好,EF-1α+Cytb组合的稳定性最佳,是玛氏骨条藻基因表达研究的理想内参基因,而其他基因的表达稳定性较差,不适合作为内参基因。本研究为玛氏骨条藻基因表达研究过程中内参基因的选择提供了方法学上的依据。
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| 关 键 词： | 铁处理 玛氏骨条藻 内参基因 实时荧光定量PCR |
| 收稿时间： | 2019-03-11 |
| 修稿时间： | 2019-04-23 |
Screening of the reference genes of Skeletonema marinoi under different concentration of Fe3+ conditions in real-time quantitative PCR analysis |
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| Zhang Mei,Xing Yongze,Zhen Yu,Mi Tiezhu and Yu Zhigang. Screening of the reference genes of Skeletonema marinoi under different concentration of Fe3+ conditions in real-time quantitative PCR analysis[J]. Acta Oceanologica Sinica (in Chinese), 2020, 42(2): 124-133. DOI: 10.3969/j.issn.0253-4193.2020.02.013 |
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| Authors: | Zhang Mei Xing Yongze Zhen Yu Mi Tiezhu Yu Zhigang |
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| Affiliation: | Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China;Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China,Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China;Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China,Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China;Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China,Key Laboratory of Marine Environment and Ecology, Ministry of Education, Ocean University of China, Qingdao 266100, China;Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;College of Environmental Science and Engineering, Ocean University of China, Qingdao 266100, China and Laboratory for Marine Ecology and Environmental Science, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;Key Laboratory of Marine Chemical Theory and Technology, Ministry of Education, Ocean University of China, Qingdao 266100, China |
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| Abstract: | Real-time quantitative PCR (qRT-PCR) is a common method for quantitative analysis of gene expression. Selection of appropriate reference genes is essential for the accurate analysis of target gene expression levels. In this study, the expression of seven reference genes of Cytb, EF-1α, HPRT, UBC, GAPDH, β-actin and β-tubulin was quantitatively analyzed with different concentrations of iron concentration. The GeNorm, NormFinder and BestKeeper software comprehensively evaluated the stability of these reference genes. The results showed that the expression stability of Cytb and EF-1α was better, and the combination of EF-1α + Cytb was the best. It could be used as a reference gene for the study of gene expression in Skeletonema marinoi, while the expression stability of other genes was poor, and they were not suitable for being used as a reference gene. This study provides a methodological basis for the selection of reference genes during the study of gene expression in S. marinoi. |
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| Keywords: | Fe3+ conditions Skeletonema marinoi reference genes real-time quantitative PCR |
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