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PMID: 12869712 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

RNA (New York, N.Y.) ·Vol. 9 ·No. 8 ·2003-08-00 ·Pages 1019-24

Harger JW, Dinman JD

Abstract

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.

MeSH Terms
Animals Base Sequence Coleoptera Frameshifting, Ribosomal Genes, Reporter Luciferases/metabolism Molecular Sequence Data Protein Biosynthesis RNA, Messenger/genetics Saccharomyces cerevisiae/genetics
Chemicals
RNA, Messenger Luciferases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Harger Jason W
Department of Molecular Genetics, Microbiology and Immunology, UMDNJ Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA.
Dinman Jonathan D
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Article Info
Journal
RNA (New York, N.Y.)
Abbr.
RNA
ISSN
1355-8382
DOI
10.1261/rna.5930803
Published
2003-08-00
Pages
1019-24
Language
English
Region
United States
NLM ID
9509184
PMCID
PMC1236998
Subset
IM
Grants
NIGMS NIH HHS · R01 GM058859 · United States
NIGMS NIH HHS · R01 GM58859 · United States
External Links
PubMed source DOI: 10.1261/rna.5930803 PMC Full Text View journal details
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