Evaluating Terminator Strength Based on Differentiating Effects on Transcription and Translation
Evaluating Terminator Strength Based on Differentiating Effects on Transcription and Translation
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基于转录和翻译差异效应评估终止子强度
DOI：
10.1002/cbic.202000068
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发表时间：
2020-07-16
期刊：
影响因子：
3.2
通讯作者：
Xu, Zhenghong
中科院分区：
文献类型：
--
作者：
He, Zhiyun;Duan, Yanting;Xu, Zhenghong
转录终止子起终止基因转录进程的作用，是基因回路中必不可少的元件。终止子主要有两个功能：终止基因转录和提高基因转录物在翻译过程中的稳定性。因此，我们考虑了终止子在基因转录和翻译中不同作用的详细特征，包括转录关闭度（α）和上游mRNA保护能力（β），以及反映终止子整体调控作用的表观终止效率（η）。基于双报告基因系统，我们构建了三个终止子探针质粒来研究大肠杆菌的每个特征。根据多元回归分析，转录关闭程度和上游mRNA保护能力对表观终止效率的贡献几乎相等。对12个终止子的序列分析表明，终止子序列以GC碱基为主，并且终止子茎结构中GC碱基的高比例可能与高度的转录关闭有关。这种终止子的全面表征进一步加深了我们对终止子在基因表达中的作用的理解，并为合成终止子的设计提供了指导。
Transcription terminators play a role in terminating the progress of gene transcription, and are thus essential elements in the gene circuit. Terminators have two main functions: terminating gene transcription and improving the stability of gene transcripts during translation. We therefore considered the detailed characteristics of terminators in relation to their different roles in gene transcription and translation, including transcription shut‐down degree (α) and upstream mRNA protection capacity (β), and apparent termination efficiency (η) reflecting the overall regulatory effect of the terminator. Based on a dual‐reporter gene system, we constructed three terminator‐probe plasmids to investigate each characteristic in Escherichia coli. According to multiple regression analysis, the transcription shut‐down degree and the upstream mRNA protection capacity contributed almost equally to the apparent termination efficiency. Sequence analysis of 12 terminators demonstrated that the terminator sequence was dominated by GC bases, and that a high ratio of GC bases in the stem structure of terminators might be associated with a high degree of transcription shut‐down. This comprehensive characterization of terminators furthers our understanding of the role of terminators in gene expression and provides a guide for synthetic terminator design.