Open AccessIn Vitro Suppression as a Tool for the Investigation of Translation Initiation
Author(s) -
Vladimir A. Karginov,
Sergey Mamaev,
Sidney M. Hecht
Publication year - 1997
Publication title -
nucleic acids research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 9.008
H-Index - 537
eISSN - 1362-4954
pISSN - 0305-1048
DOI - 10.1093/nar/25.19.3912
Subject(s) - biology , reticulocyte , transfer rna , translation (biology) , protein biosynthesis , escherichia coli , in vitro , messenger rna , nonsense mediated decay , eukaryotic translation , microbiology and biotechnology , biochemistry , rna , gene , rna splicing
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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