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

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

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 ce...

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Veröffentlicht in:RNA (Cambridge) 2003-08, Vol.9 (8), p.1019-1024
Hauptverfasser: Harger, Jason W, Dinman, Jonathan D
Format: Artikel
Sprache:eng
Schlagworte:
Animals
Base Sequence
Coleoptera
Frameshifting, Ribosomal
Genes, Reporter
Luciferases - metabolism
Method
Molecular Sequence Data
Protein Biosynthesis
RNA, Messenger - genetics
Saccharomyces cerevisiae - genetics
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creator Harger, Jason W
Dinman, Jonathan D
description 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.
doi_str_mv 10.1261/rna.5930803
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subjects Animals
Base Sequence
Coleoptera
Frameshifting, Ribosomal
Genes, Reporter
Luciferases - metabolism
Method
Molecular Sequence Data
Protein Biosynthesis
RNA, Messenger - genetics
Saccharomyces cerevisiae - genetics
title An in vivo dual-luciferase assay system for studying translational recoding in the yeast Saccharomyces cerevisiae
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