In vitro suppression as a tool for the investigation of translation initiation

Vladimir A. Karginov; Sergey V. Mamaev; Sidney M. Hecht

doi:10.1093/nar/25.19.3912

In vitro suppression as a tool for the investigation of translation initiation

Vladimir A. Karginov, Sergey V. Mamaev, Sidney M. Hecht

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

Abstract

An in vitro protein synthesizing system that employs rabbit reticulocyte lysates has been employed for protein production from mRNAs containing nonsense (UAG) codons in the presence of misacylated suppressor tRNAs. The system includes a misacylated Escherichia coli tRNA(Ala)(CUA) that functions at least as efficiently as any suppressor tRNA transcript reported to date and which has been shown not to be a substrate for (re)activation by alanyl-tRNA synthetase. Application of the optimized system for preparation of dihydrofolate analogs has also permitted analysis of competing mechanisms that control the sites(s) of translation initiation.

Original language	English (US)
Pages (from-to)	3912-3916
Number of pages	5
Journal	Nucleic acids research
Volume	25
Issue number	19
DOIs	https://doi.org/10.1093/nar/25.19.3912
State	Published - Oct 1 1997
Externally published	Yes

ASJC Scopus subject areas

Genetics

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title = "In vitro suppression as a tool for the investigation of translation initiation",

abstract = "An in vitro protein synthesizing system that employs rabbit reticulocyte lysates has been employed for protein production from mRNAs containing nonsense (UAG) codons in the presence of misacylated suppressor tRNAs. The system includes a misacylated Escherichia coli tRNA(Ala)(CUA) that functions at least as efficiently as any suppressor tRNA transcript reported to date and which has been shown not to be a substrate for (re)activation by alanyl-tRNA synthetase. Application of the optimized system for preparation of dihydrofolate analogs has also permitted analysis of competing mechanisms that control the sites(s) of translation initiation.",

author = "Karginov, {Vladimir A.} and Mamaev, {Sergey V.} and Hecht, {Sidney M.}",

note = "Funding Information: We thank Drs John Abelson and George Komatsoulis (California Institute of Technology) for the plasmid containing DNAfol and Drs Paul Schimmel (Massachusetts Institute of Technology) and Ya-Ming Hou (Thomas Jefferson University) for the plasmid that permitted us to elaborate E.coli tRNACUAAla (–CA). This work was supported by NIH Research Grant GM43328.",

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AU - Mamaev, Sergey V.

AU - Hecht, Sidney M.

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In vitro suppression as a tool for the investigation of translation initiation

Abstract

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