Schaeffer Daneen, Meaux Stacie, Clark Amanda, van Hoof Ambro
University of Texas Health Science Center-Houston, Department of Microbiology and Molecular Genetics, Houston, Texas, USA.
Methods Enzymol. 2008;448:227-39. doi: 10.1016/S0076-6879(08)02612-8.
A 3'-exoribonuclease complex, termed the exosome, has important functions in the cytoplasm, as well as in the nucleus, and is involved in 3'-processing and/or decay of many RNAs. This chapter will discuss methods to study cytoplasmic exosome function in yeast with in vivo approaches. The first section will describe mutants that are available to study the processing or decay of a specific RNA by the nuclear or cytoplasmic exosome. The second section will discuss methods to determine whether the cytoplasmic exosome is functional under a specific condition(s) with reporter mRNAs that are known substrates of this complex.
一种名为外切体的3'-外切核糖核酸酶复合体在细胞质以及细胞核中都具有重要功能，并且参与许多RNA的3'-加工和/或降解过程。本章将讨论利用体内方法研究酵母细胞质中外切体功能的方法。第一部分将描述可用于研究核内或细胞质外切体对特定RNA进行加工或降解的突变体。第二部分将讨论使用已知为该复合体底物的报告mRNA来确定细胞质外切体在特定条件下是否具有功能的方法。
