Enhancement of translation initiation by A/T-Rich sequences downstream of the initiation codon in Escherichia coli
Enhancement of translation initiation by A/T-Rich sequences downstream of the initiation codon in Escherichia coli
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DOI：
10.1159/000077244
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发表时间：
2003-01-01
影响因子：
1.2
通讯作者：
Inouye, M
中科院分区：
文献类型：
--
作者：
Qing, GL;Xia, B;Inouye, M
起始密码子下游的区域构成了翻译起始信号的一部分，显著影响着大肠杆菌中蛋白的表达水平。为了确定其对翻译起始的影响，我们在lacZ基因起始密码子下游随机插入12个碱基序列。共分离出119个随机克隆，显示出比对照lacZ基因更高的β -半乳糖苷酶活性，并对其进行测序。对这些克隆的分析表明，它们的插入序列在A和T上明显丰富，而在G上却很少，它们之间没有一致的序列。足印实验和多聚体分析证实，富含A/ t的序列在起始水平上促进了翻译。总的来说，目前的数据表明，起始密码子后区域的A/T丰富度在大肠杆菌基因表达中起着重要作用。版权所有(C) 2003 S. KargerAG，巴塞尔。
The region located downstream of the initiation codon constitutes part of the translation initiation signal, significantly affecting the level of protein expression in E. coli. In order to determine its influence on translation initiation, we inserted random 12-base sequences downstream of the initiation codon of the lacZ gene. A total of 119 random clones showing higher beta-galactosidase activities than the control lacZ gene were isolated and subsequently sequenced. Analysis of these clones revealed that their insertion sequences are strikingly rich in A and T, but poor in G, with no consensus sequences among them. Toeprinting experiments and polysome profile analysis confirmed that the A/T-rich sequences enhance translation at the level of initiation. Collectively, the present data demonstrate that A/T richness of the region following the initiation codon plays a significant role in E. coli gene expression. Copyright (C) 2003 S. KargerAG, Basel.