Development of a tRNA-dependent in vitro translation system

被引：19

作者：

Jackson, RJ

Napthine, S

Brierley, I

机构：

[1] Univ Cambridge, Dept Biochem, Cambridge CB2 1GA, England

[2] Univ Cambridge, Dept Pathol, Div Virol, Cambridge CB2 1QP, England

来源：

RNA | 2001年 / 7卷 / 05期

基金：

英国医学研究理事会;

关键词：

leaky termination; programmed ribosomal frame-shifting; rabbit reticulocyte lysate; wheat germ; system;

D O I：

10.1017/S1355838201002539

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：

A method is described for depleting rabbit reticulocyte lysates and wheat germ extracts of endogenous tRNAs by affinity chromatography using a matrix generated by coupling ethanolamine to epoxy-activated Sepharose 6B. Greater than 90% depletion of tRNA is achieved with the result that translation becomes in effect absolutely dependent on added tRNA. This depletion procedure should prove very useful for studying the influence of tRNA concentration, and the spectrum of the tRNA population, on recoding events such as programmed frameshifting and readthrough of termination codons.

引用

页码：765 / 773

页数：9

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