Dual-luciferase reporter system for studying recoding signals

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Publication Type Journal Article
School or College School of Medicine; College of Science; College of Engineering
Department Human Genetics; Howard Hughes Medical Institute; Bioengineering; Biology
Creator Gesteland, Raymond F.; Atkins, John F.; Ingram, Jennifer A.; Kelly, Paul J.; Grentzmann, Guido
Title Dual-luciferase reporter system for studying recoding signals
Date 1998
Description A new reporter system has been developed for measuring translation coupling efficiency of recoding mechanisms such as frameshifting or readthrough. A recoding test sequence is cloned in between the renilla and firefly luciferase reporter genes and the two luciferase activities are subsequently measured in the same tube. The normalized ratio of the two activities is proportional to the efficiency with which the ribosome "reads" the recoding signal making the transition from one open reading frame to the next. The internal control from measuring both activities provides a convenient and reliable assay of efficiency. This is the first enzymatic dual reporter assay suitable for in vitro translation. Translation signals can be tested in vivo and in vitro from a single construct, which allows an intimate comparison between the two systems. The assay is applicable for high throughput screening procedures. The dual-luciferase reporter system has been applied to in vivo and in vitro recoding of HIV-1 gag-pol, MMTV gag-pro, MuLV gag-pol, and human antizyme.
Type Text
Publisher Cold Spring Harbor Lab Press
First Page 479
Last Page 486
Subject Amino Acid Sequence; Genes, Reporter; HIV; Antizyme; Translation
Subject MESH Base Sequence; Molecular Sequence Data; Protein Biosynthesis; Reading Frames
Language eng
Bibliographic Citation Grentzmann, G., Ingram, J.A., Kelly, P.J., Gesteland, R.F., and Atkins, J.F. 1998. A dual-luciferase reporter system for studying recoding signals. RNA 4:479-486
Rights Management © Cold Spring Harbor Laboratory Press-CSHL Press
Format Medium application/pdf
Identifier ir-main,12747
ARK ark:/87278/s6hm5t1n
Setname ir_uspace
ID 706537
Reference URL https://collections.lib.utah.edu/ark:/87278/s6hm5t1n
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