确定mRNA是否为酿酒酵母中无义介导的mRNA降解的底物。

Determining if an mRNA is a Substrate of Nonsense-Mediated mRNA Decay in Saccharomyces cerevisiae.

作者信息

Johansson Marcus J O

机构信息

Department of Molecular Biology, Umeå University, 901 87, Umeå, Sweden.

出版信息

Methods Mol Biol. 2017;1507:169-177. doi: 10.1007/978-1-4939-6518-2_13.

DOI:10.1007/978-1-4939-6518-2_13

PMID:27832540

Abstract

Nonsense-mediated mRNA decay (NMD) is a conserved eukaryotic quality control mechanism which triggers decay of mRNAs harboring premature translation termination codons. In this chapter, I describe methods for monitoring the influence of NMD on mRNA abundance and decay rates in Saccharomyces cerevisiae. The descriptions include detailed methods for growing yeast cells, total RNA isolation, and Northern blotting. Although the chapter focuses on NMD, the methods can be easily adapted to assess the effect of other mRNA decay pathways.

摘要

无义介导的mRNA降解（NMD）是一种保守的真核生物质量控制机制，它会触发携带提前翻译终止密码子的mRNA的降解。在本章中，我描述了监测NMD对酿酒酵母中mRNA丰度和降解速率影响的方法。这些描述包括培养酵母细胞、总RNA分离和Northern印迹的详细方法。尽管本章重点关注NMD，但这些方法可以很容易地适用于评估其他mRNA降解途径的作用。