Translational coupling at the intercistronic boundary of an artificially constructed operon in Escherichia coli.

G Szabó, P Venetianer
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Abstract

By using recombinant DNA techniques an artificial operon was constructed that codes for two fusion proteins under the control of the beta-lactamase promoter of plasmid pBR322. The two proteins are: 1. beta-lactamase-trp B (43 kd) 2. trpA-beta-galactosidase (120 kd). Frameshift mutations in the N-terminal region of the first gene resulted in a dramatic reduction in the synthesis of the protein coded by the second gene. This strong polar effect could not be accounted for by correspondingly lower level of the distal region of the messenger RNA, only by "translational coupling" due to the overlap of the termination codon of the first gene with the initiation codon of the second gene. It was concluded that the strong "translational coupling" observed in this artificial operon can be generally used to ensure coordinated high-level synthesis of proteins in operons constructed by recombinant DNA techniques.

大肠杆菌中人工构建的操纵子在顺子间边界的翻译耦合。
利用重组DNA技术,在pBR322质粒β -内酰胺酶启动子的控制下,构建了一个编码两个融合蛋白的人工操纵子。这两种蛋白质是:1;β -内酰胺酶-trp B (43 kd)trpa - β -半乳糖苷酶(120kd)。第一个基因n端区域的移码突变导致第二个基因编码的蛋白质合成急剧减少。这种强烈的极性效应不能用信使RNA远端相应较低的水平来解释,只能用第一个基因的终止密码子与第二个基因的起始密码子重叠而产生的“翻译偶联”来解释。由此可见,该人工操纵子具有很强的“翻译偶联性”,可用于保证重组DNA技术构建的操纵子中蛋白质的协调高水平合成。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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