Khoury Marie P, Marcel Virginie, Fernandes Kenneth, Diot Alexandra, Lane David P, Bourdon Jean-Christophe
CR-UK Cell Transformation Research Group, Centre for Oncology and Molecular Medicine, Ninewells Hospital, University of Dundee, Dundee, UK.
Methods Mol Biol. 2013;962:1-14. doi: 10.1007/978-1-62703-236-0_1.
The TP53 gene expresses at least nine different mRNA variants (p53 isoform mRNAs), including the one encoding the canonical p53 tumor suppressor protein. We have developed scientific tools to specifically detect and quantify p53 isoform expression at mRNA level by nested RT-PCR (reverse transcription-polymerase chain reaction) and quantitative real-time RT-PCR (RT-qPCR using the TaqMan(®) chemistry). Here, we describe these two methods, while highlighting essential points with regard to the analysis of p53 isoform mRNA expression.
TP53基因至少表达九种不同的mRNA变体（p53亚型mRNA），包括编码经典p53肿瘤抑制蛋白的变体。我们开发了科学工具，通过巢式逆转录-聚合酶链反应（RT-PCR）和定量实时RT-PCR（使用TaqMan®化学方法的RT-qPCR）在mRNA水平特异性检测和定量p53亚型表达。在此，我们描述这两种方法，同时强调p53亚型mRNA表达分析的要点。
