An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae

被引：131

作者：

Harger, JW

Dinman, JD

机构：

[1] Univ Maryland, Dept Mol Genet & Cell Biol, College Pk, MD 20742 USA

[2] Univ Med & Dent New Jersey, Robert Wood Johnson Med Sch, Dept Mol Genet Microbiol & Immunol, Piscataway, NJ 08854 USA

来源：

RNA | 2003年 / 9卷 / 08期

关键词：

virus; ribosome; translation; frameshifting; bicistronic; lysate;

D O I：

10.1261/rna.5930803

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A new in vivo assay system has been developed to study programmed frameshifting in the yeast Saccharomyces cerevisiae. Frameshift signals are inserted between the Renilla and firefly luciferase reporter genes contained in a yeast expression vector and the two activities are directly measured from cell lysates in one tube. Similar to other bicistronic reporter systems, this one allows the efficient estimation of recoding efficiency by comparison of the normalized activity ratios from each luciferase protein. The assay system has been applied to HIV-1 and L-A directed programmed -1 frameshifting and Ty1 and Ty3 directed +1 frameshifting. The assay system is amenable to high-throughput screening.

引用

页码：1019 / 1024

页数：6

共 21 条

[1]

BECKETT GJ, 1941, P NATL ADAC SCI, V27, P499

[2] Nonsense-mediated decay mutants do not affect programmed-1 frameshifting [J].