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

被引:19
作者
Karginov, VA
Mamaev, SV
Hecht, SM
机构
[1] UNIV VIRGINIA,DEPT CHEM,CHARLOTTESVILLE,VA 22901
[2] UNIV VIRGINIA,DEPT BIOL,CHARLOTTESVILLE,VA 22901
关键词
D O I
10.1093/nar/25.19.3912
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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(CUA)(Ala) 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.
引用
收藏
页码:3912 / 3916
页数:5
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