一种依赖于转运RNA的体外翻译系统的开发。
Development of a tRNA-dependent in vitro translation system.
作者信息
Jackson R J, Napthine S, Brierley I
机构信息
Department of Biochemistry, University of Cambridge, United Kingdom.
出版信息
RNA. 2001 May;7(5):765-73. doi: 10.1017/s1355838201002539.
Abstract
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.
摘要
本文描述了一种通过亲和色谱法去除兔网织红细胞裂解物和小麦胚芽提取物中内源性tRNA的方法，该方法使用通过将乙醇胺偶联到环氧活化的琼脂糖凝胶6B上生成的基质。tRNA的去除率超过90%，结果是翻译实际上完全依赖于添加的tRNA。这种去除程序对于研究tRNA浓度和tRNA群体谱对重编码事件（如程序性移码和终止密码子通读）的影响应该非常有用。
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