Yang Yanfang, Hou Shuang, Cui Guanghong, Chen Shilin, Wei Jianhe, Huang Luqi
Institute of Medicinal Plant Development, Chinese Academy of Medicinal Sciences and Peking Union Medical College, Beijing, China.
Mol Biol Rep. 2010 Jan;37(1):507-13. doi: 10.1007/s11033-009-9703-3. Epub 2009 Aug 13.
Five reference genes, 18S, EF1alpha, alpha-Tubulin, Ubiquitin and Actin, from Salvia miltiorrhiza were analyzed as internal controls for gene expression profiling assay using quantitative real-time polymerase chain reaction (qRT-PCR). The five candidate genes were measured for their transcriptional level in seven tissues, including roots, stems, leaves, sepals, petals, stamens and pistils. Then they were ranked by the GeNorm tool. The results showed that Actin and Ubiquitin were the most stable whereas EF1alpha and 18S did not favor normalization of qRT-PCR results in these tissues. Expression levels of the SmDXR gene were studied in parallel, with Actin and Ubiquitin both or each as reference in the seven tissues, and varying relative quantifications of the SmDXR gene in seven tissues. This study indicated that selection of the most stable genes plays an important role in gene expression profiling assays.
对丹参的五个参考基因18S、EF1α、α-微管蛋白、泛素和肌动蛋白进行分析，作为使用定量实时聚合酶链反应（qRT-PCR）进行基因表达谱分析的内参。测定了这五个候选基因在根、茎、叶、萼片、花瓣、雄蕊和雌蕊七种组织中的转录水平。然后用GeNorm工具对它们进行排名。结果表明，肌动蛋白和泛素是最稳定的，而EF1α和18S不利于这些组织中qRT-PCR结果的标准化。同时研究了SmDXR基因在七种组织中的表达水平，分别以肌动蛋白和泛素作为参考，结果显示SmDXR基因在七种组织中的相对定量有所不同。本研究表明，选择最稳定的基因在基因表达谱分析中起着重要作用。
