Methods for studying IRES-mediated translation of positive-strand RNA viruses

被引：17
作者：
Wang, Qing S. [1 ]
Au, Hilda H. T. [1 ]
Jan, Eric [1 ]
机构：
[1] Univ British Columbia, Dept Biochem & Mol Biol, Vancouver, BC V6T 1Z3, Canada
来源：
METHODS | 2013年 / 59卷 / 02期
基金：
加拿大自然科学与工程研究理事会;
关键词：
RNA; Ribosome; 2A peptide; Toeprinting; Bicistronic reporter; RIBOSOME ENTRY SITE; CRICKET PARALYSIS VIRUS; EUKARYOTIC MESSENGER-RNAS; HOST PROTEIN-SYNTHESIS; HEPATITIS-C VIRUS; INTERNAL RIBOSOME; IN-VITRO; INDEPENDENT TRANSLATION; INITIATION-FACTORS; CODING SEQUENCE;
D O I：
10.1016/j.ymeth.2012.09.004
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Internal ribosome entry sites are RNA elements that mediate translation in a cap-independent manner. A subset of positive strand RNA viruses utilize an IRES mechanism as a viral strategy to ensure efficient viral protein synthesis. IRES elements vary in sequence, structure, and factor requirements between virus families. Here, we describe methods to determine IRES activity and approaches to study the regulation and function of IRES-mediated translation both in vitro and in vivo. Finally, we describe a new IRES-directed reporter system which exploits the 2A 'self-cleavage' or 'stop-go' peptide for optimal detection of IRES activity. (C) 2012 Elsevier Inc. All rights reserved.
引用
收藏
页码：167 / 179
页数：13
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