通过对mRNA水平进行定量来鉴定基因表达的变化。

Identification of changes in gene expression by quantitation of mRNA levels.

作者信息

Olivas Wendy M

机构信息

Department of Biology, University of Missouri - St. Louis, St. Louis, MO, USA.

出版信息

Methods Mol Biol. 2008;419:243-58. doi: 10.1007/978-1-59745-033-1_17.

DOI:10.1007/978-1-59745-033-1_17

PMID:18369988

Abstract

Sequence elements within mRNA-untranslated regions and their binding partners are key controllers of mRNA stability. Changes in mRNA stability can often be detected by changes in steady-state mRNA abundance, or a more careful analysis of mRNA half-lives can be performed following transcriptional repression. This chapter presents methods to isolate RNA from both yeast and mammalian cells for either steady-state or half-life analyses. In addition, two reliable methods to quantitate mRNA levels, northern blot analysis and real-time PCR, are outlined and compared.

摘要

信使核糖核酸（mRNA）非翻译区的序列元件及其结合伴侣是mRNA稳定性的关键调控因素。mRNA稳定性的变化通常可通过稳态mRNA丰度的变化来检测，或者在转录抑制后对mRNA半衰期进行更细致的分析。本章介绍了从酵母细胞和哺乳动物细胞中分离RNA以进行稳态分析或半衰期分析的方法。此外，还概述并比较了两种可靠的定量mRNA水平的方法，即Northern印迹分析和实时聚合酶链反应（PCR）。