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Nucleic Acids Research, Vol 25, Issue 19 3912-3916, Copyright © 1997 by Oxford University Press

ARTICLES

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

VA Karginov, SV Mamaev and SM Hecht
Departments of Chemistry and Biology, University of Virginia, Charlottesville, VA 22901, USA.

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 tRNAAlaCUA 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.
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