Tokunaga Kazuaki, Tani Tokio
Department of Biological Sciences, Graduate School of Science and Technology, Kumamoto University, Kurokami, Kumamoto, Japan.
Curr Protoc Cell Biol. 2008 Dec;Chapter 22:Unit 22.13. doi: 10.1002/0471143030.cb2213s41.
Transport of mRNA from the nucleus to the cytoplasm is an essential process for gene expression in eukaryotic cells. In this unit, methods for monitoring nuclear mRNA export are described. Visualization of cellular mRNAs by fluorescence in situ hybridization with oligo(dT) probes is effectively applied to monitoring mRNA export from the nucleus in yeast and mammalian cells. In addition to the protocols for fluorescence in situ hybridization, this unit includes an alternate method that the authors have been developing for visual analysis of nuclear mRNA export in living mammalian cells by microinjection of fluorescently labeled pre-mRNA into the nuclei.
信使核糖核酸(mRNA)从细胞核转运至细胞质是真核细胞中基因表达的一个重要过程。在本单元中,将描述监测细胞核mRNA输出的方法。通过与寡聚(dT)探针进行荧光原位杂交来可视化细胞mRNA,可有效地用于监测酵母和哺乳动物细胞中mRNA从细胞核的输出。除了荧光原位杂交的实验方案外,本单元还包括一种替代方法,这是作者一直在开发的,通过将荧光标记的前体mRNA显微注射到活的哺乳动物细胞核中,对细胞核mRNA输出进行视觉分析。