通过米勒染色质铺展法对酿酒酵母中RNA转录和加工进行电子显微镜观察。

Electron microscope visualization of RNA transcription and processing in Saccharomyces cerevisiae by Miller chromatin spreading.

作者信息

Osheim Yvonne N, French Sarah L, Sikes Martha L, Beyer Ann L

机构信息

Department of Microbiology, University of Virginia Health System, Charlottesville, VA, USA.

出版信息

Methods Mol Biol. 2009;464:55-69. doi: 10.1007/978-1-60327-461-6_4.

DOI:10.1007/978-1-60327-461-6_4

PMID:18951179

Abstract

The Miller chromatin spreading technique for electron microscopic visualization of gently dispersed interphase chromatin has proven extremely valuable for analysis of genetic activities in vivo. It provides a unique view of transcription and RNA processing at the level of individual active genes. The budding yeast Saccharomyces cerevisiae has also been an invaluable model system for geneticists and molecular biologists. In this chapter, we describe methods for applying the Miller chromatin-spreading method to Saccharomyces cerevisiae. This allows one to use electron microscopic visualization of a gene of interest to study effects of specific mutations on gene activity. We are applying the method to study transcription and processing of ribosomal RNA.

摘要

用于电子显微镜观察温和分散的间期染色质的米勒染色质铺展技术，已被证明对体内遗传活动的分析极具价值。它在单个活性基因水平上提供了转录和RNA加工的独特视角。芽殖酵母酿酒酵母对遗传学家和分子生物学家来说也是一个非常有价值的模型系统。在本章中，我们描述了将米勒染色质铺展方法应用于酿酒酵母的方法。这使得人们能够利用感兴趣基因的电子显微镜观察来研究特定突变对基因活性的影响。我们正在应用该方法研究核糖体RNA的转录和加工。