Pellegrini Olivier, Mathy Nathalie, Condon Ciarán, Bénard Lionel
CNRS UPR 9073 (affiliated with Université de Paris 7 - Denis Diderot), Institut de Biologie Physico-Chimique, Paris, France.
Methods Enzymol. 2008;448:167-83. doi: 10.1016/S0076-6879(08)02609-8.
The major cytoplasmic 5' to 3'-exoribonuclease activity is carried out by the Xrn1 protein in eukaryotic cells. A number of different approaches can be used to study multifunctional Xrn1 protein activity in vitro. In this chapter, we concentrate on methods used in our laboratory to analyze Xrn1 5' to 3'-exoribonuclease activity. Some of these techniques may also be suitable for detecting 3' to 5'-exoribonuclease or endoribonuclease activity. For these reasons, these assays can be used to isolate new proteins with ribonuclease activity and, when performed in combination with in vivo experiments, will contribute to a new level of understanding of the function of these factors.
真核细胞中主要的胞质5'至3'外切核糖核酸酶活性由Xrn1蛋白执行。可以使用多种不同方法在体外研究多功能Xrn1蛋白活性。在本章中，我们专注于我们实验室用于分析Xrn1 5'至3'外切核糖核酸酶活性的方法。其中一些技术也可能适用于检测3'至5'外切核糖核酸酶或内切核糖核酸酶活性。基于这些原因，这些测定法可用于分离具有核糖核酸酶活性的新蛋白质，并且当与体内实验结合进行时，将有助于对这些因子的功能有新的理解。
