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基于衰减的双荧光蛋白报告基因用于筛选翻译抑制剂。

Attenuation-based dual-fluorescent-protein reporter for screening translation inhibitors.

机构信息

Department of Chemistry and A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia.

出版信息

Antimicrob Agents Chemother. 2012 Apr;56(4):1774-83. doi: 10.1128/AAC.05395-11. Epub 2012 Jan 17.

Abstract

A reporter construct was created on the basis of the transcription attenuator region of the Escherichia coli tryptophan operon. Dual-fluorescent-protein genes for red fluorescent protein and cerulean fluorescent protein were used as a sensor and internal control of gene expression. The sequence of the attenuator was modified to avoid tryptophan sensitivity while preserving sensitivity to ribosome stalling. Antimicrobial compounds which cause translation arrest at the stage of elongation induce the reporter both in liquid culture and on an agar plate. This reporter could be used for high-throughput screening of translation inhibitors.

摘要

一个记者构建了基于转录衰减器区域的大肠杆菌色氨酸操纵子。双荧光蛋白基因的红色荧光蛋白和青色荧光蛋白作为传感器和基因表达的内部对照。衰减序列进行了修饰,在保留对核糖体停滞敏感性的同时避免色氨酸敏感性。引起延伸阶段翻译停滞的抗菌化合物在液体培养和琼脂平板上诱导报告基因。该报告基因可用于高通量筛选翻译抑制剂。

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