Selection of candidate housekeeping controls in tomato plants using EST data
Selection of candidate housekeeping controls in tomato plants using EST data
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DOI：
10.2144/03354st04
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发表时间：
2003-10-01
期刊：
影响因子：
2.7
通讯作者：
Davies, E
中科院分区：
文献类型：
--
作者：
Coker, JS;Davies, E
因为管家基因的表达水平在大多数组织中相对恒定，所以当定量基因表达时，它们通常是有用的对照。我们提出了一种分析方法，用于确定候选管家控制使用表达序列标签（EST）从基因组研究所番茄基因索引。我们发现了127个转录本的相对表达水平，并计算了表达水平（对于给定的转录本）在2倍至10倍范围内的cDNA文库的百分比。当将所有文库一起考虑时，最高等级的管家对照包括DnaJ样蛋白质的转录物、抑制性控制的肿瘤蛋白质、两种α-微管蛋白、亲环蛋白和甘油醛-3-磷酸脱氢酶（GAPDH）。对于每个器官（叶根、果实和花），发现至少一种转录物在所有相应文库中的2倍表达范围内发生。这些包括α-微管蛋白、DnaJ样蛋白、磷酸甘油酸激酶和GAPDH的转录物，尽管不同的转录物似乎比其他转录物更适合于不同的组织。该分析方法可用于鉴定特定组织中和特定表达水平下的候选管家对照，并且与存在重要EST数据的任何物种相关。
Because the expression levels of housekeeping genes art, relatively constant in most tissues, they, are often useful controls when quantifying gene expression. We present an analytical method for identifying candidate housekeeping controls using expressed sequence tags (ESTs) from The Institute for Genomic Research Tomato Gene Index. We found relative expression levels for a collection of 127 transcripts and calculated the percentage of cDNA libraries that had expression levels (for a given transcript) within 2-fold through 10-fold ranges. When all libraries were considered together, the highest ranked housekeeping controls included transcripts for a DnaJ-like protein, translationally controlled tumor protein, two alpha-tubulins, cyclophilin, and glyceraldehyde-3-phosphate dehydrogenase (GAPDH). For each organ (leaf root, fruit, and flower), at least one transcript was found that occurred within a 2-fold range of expression in all respective libraries. These included transcripts for alpha-tubulin, DnaJ-like proteins, phospohoglycerate kinase, and GAPDH, although different transcripts appear better suited than others for different tissues. This analytical method is useful for identifying candidate housekeeping controls in particular tissues and at particular levels of expression and would be relevant for any species for which significant EST data exist.