Pachot Alexandre, Blond Jean-Luc, Mougin Bruno, Miossec Pierre
Department of Human Genetics, BioMérieux, 69280 Marcy-l'Etoile, France.
J Biotechnol. 2004 Oct 19;114(1-2):121-4. doi: 10.1016/j.jbiotec.2004.07.001.
Quantitative real-time RT-PCR is a very powerful technique for measuring gene expression at the mRNA level. In order to compare mRNA expression in different experimental or clinical conditions, expression of a target gene has to be normalized to an appropriate internal standard, which is generally a housekeeping gene. In our study, we have tested several housekeeping genes in peripheral whole blood of healthy volunteers and patients suffering from inflammatory diseases. A first analysis of 91 samples illustrated that the mRNA expression of peptidylpropyl isomerase B (PPIB) encoding for cyclophilin B protein, is more stable than beta actin and glyceraldehyde-3-phosphate dehydrogenase, which are both commonly selected as internal standard. Among the three genes tested, beta actin displayed the highest inter-sample variation of expression. The constancy of PPIB mRNA expression was further confirmed by 214 additional samples. In conclusion, we showed that PPIB, in contrast to beta actin and glyceraldehyde-3-phosphate dehydrogenase, is a suitable housekeeping gene in human peripheral blood.
定量实时逆转录聚合酶链反应（Quantitative real-time RT-PCR）是一种在mRNA水平测量基因表达的非常强大的技术。为了比较不同实验或临床条件下的mRNA表达，靶基因的表达必须用合适的内参进行标准化，内参通常是一个管家基因。在我们的研究中，我们检测了健康志愿者和患有炎症性疾病患者外周全血中的几个管家基因。对91个样本的初步分析表明，编码亲环素B蛋白的肽基脯氨酰异构酶B（PPIB）的mRNA表达比β-肌动蛋白和甘油醛-3-磷酸脱氢酶更稳定，后两者通常被选作内参。在所检测的三个基因中，β-肌动蛋白的样本间表达差异最大。另外214个样本进一步证实了PPIB mRNA表达的稳定性。总之，我们表明，与β-肌动蛋白和甘油醛-3-磷酸脱氢酶不同，PPIB是人类外周血中合适的管家基因。
