坛紫菜（红藻门）不同生活史阶段表达研究中内参基因的选择(1)。
SELECTION OF INTERNAL CONTROL GENE FOR EXPRESSION STUDIES IN PORPHYRA HAITANENSIS (RHODOPHYTA) AT DIFFERENT LIFE-HISTORY STAGES(1).
作者信息
Wu Xiaojie, Niu Jianfeng, Huang Aiyou, Xu Meiling, Wang Guangce
机构信息
College of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, ChinaKey Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences (IOCAS), Qingdao 266071, China Key Laboratory of Experimental Marine Biology, Institute of Oceanology, The Chinese Academy of Sciences (IOCAS), Qingdao 266071, China Graduate University of the Chinese Academy of Sciences, Beijing 100049, ChinaCollege of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China Key Laboratory of Experimental Marine Biology, Institute of Oceanology, the Chinese Academy of Sciences (IOCAS), Qingdao 266071, China College of Marine Science and Engineering, Tianjin University of Science and Technology, Tianjin 300457, China.
出版信息
J Phycol. 2012 Aug;48(4):1040-4. doi: 10.1111/j.1529-8817.2012.01188.x. Epub 2012 May 31.
Accurate gene quantification depends on the use of an appropriate internal control gene, which should be verified before its use for normalizing data. Housekeeping genes, which are expressed at relatively constant levels, are generally regarded as candidate internal control genes. To determine the ideal internal control for gene expression profiles for Porphyra haitanensis T. J. Chang et B. F. Zheng (Bangiales, Rhodophyta) at different life-history stages, we used absolute quantification to assess the expression levels of six housekeeping genes (18S ribosomal RNA, 30S ribosomal protein, glyceraldehyde-3-phosphate dehydrogenase, elongation factor 3, alpha-tubulin, and beta-tubulin) at the sporophyte and gametophyte stages. Housekeeping genes were selected by comparing the differences of observed copy numbers in sporophytes and in gametophytes. TubB (beta-tubulin) was found to be the optimal internal control gene, because it showed the smallest difference of gene expression. Compared with TubB, other housekeeping genes had greater variation of expression to different degrees.
准确的基因定量依赖于使用合适的内参基因，在将其用于数据标准化之前应进行验证。看家基因通常被视为候选内参基因，其表达水平相对恒定。为了确定不同生活史阶段的坛紫菜（红藻门，红毛菜纲）基因表达谱的理想内参基因，我们采用绝对定量法评估了六个看家基因（18S核糖体RNA、30S核糖体蛋白、甘油醛-3-磷酸脱氢酶、延伸因子3、α-微管蛋白和β-微管蛋白）在孢子体和配子体阶段的表达水平。通过比较孢子体和配子体中观察到的拷贝数差异来选择看家基因。发现TubB（β-微管蛋白）是最佳内参基因，因为它的基因表达差异最小。与TubB相比，其他看家基因的表达有不同程度的更大变化。
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