Jain Mukesh, Nijhawan Aashima, Tyagi Akhilesh K, Khurana Jitendra P
Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110 021, India.
Biochem Biophys Res Commun. 2006 Jun 30;345(2):646-51. doi: 10.1016/j.bbrc.2006.04.140. Epub 2006 May 3.
For accurate and reliable gene expression results, normalization of real-time PCR data is required against a control gene, which displays highly uniform expression in living organisms during various phases of development and under different environmental conditions. We assessed the gene expression of 10 frequently used housekeeping genes, including 18S rRNA, 25S rRNA, UBC, UBQ5, UBQ10, ACT11, GAPDH, eEF-1alpha, eIF-4a, and beta-TUB, in a diverse set of 25 rice samples. Their expression varied considerably in different tissue samples analyzed. The expression of UBQ5 and eEF-1alpha was most stable across all the tissue samples examined. However, 18S and 25S rRNA exhibited most stable expression in plants grown under various environmental conditions. Also, a set of two genes was found to be better as control for normalization of the data. The expression of these genes (with more uniform expression) can be used for normalization of real-time PCR results for gene expression studies in a wide variety of samples in rice.
为了获得准确可靠的基因表达结果，实时定量PCR数据需要根据一个对照基因进行标准化，该对照基因在生物体发育的各个阶段以及不同环境条件下都表现出高度一致的表达。我们评估了10个常用持家基因的基因表达，包括18S rRNA、25S rRNA、泛素结合酶（UBC）、泛素5（UBQ5）、泛素10（UBQ10）、肌动蛋白11（ACT11）、甘油醛-3-磷酸脱氢酶（GAPDH）、真核生物延伸因子-1α（eEF-1α）、真核生物翻译起始因子4A（eIF-4a）和β-微管蛋白（β-TUB），这些基因来自25个不同的水稻样本。在分析的不同组织样本中，它们的表达差异很大。在所有检测的组织样本中，UBQ5和eEF-1α的表达最稳定。然而，18S和25S rRNA在不同环境条件下生长的植物中表现出最稳定的表达。此外，还发现一组两个基因作为数据标准化的对照更好。这些基因（表达更均匀）的表达可用于水稻各种样本基因表达研究的实时定量PCR结果的标准化。
