Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species.
Evaluation and Comparison of the Efficiency of Transcription Terminators in Different Cyanobacterial Species.
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DOI：
10.3389/fmicb.2020.624011
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发表时间：
2020
影响因子：
5.2
通讯作者：
McCormick AJ
中科院分区：
文献类型：
--
作者：
Gale GAR;Wang B;McCormick AJ
蓝细菌利用阳光将二氧化碳转化为多种次生代谢物，在绿色生物技术应用方面显示出巨大的潜力。尽管蓝藻合成生物学不如其他异养模型生物成熟，但现在有一系列分子工具可用于调节和控制基因表达。基因调控仍然落后于其他模式生物的一个领域是基因转录的调节，特别是转录终止。现在异养生物中存在大量的内在转录终止子，但在蓝细菌中只研究了一小部分。随着人工基因表达系统变得更大、更复杂，短片段 DNA 含有强启动子和多个基因表达盒，有效停止转录并隔离下游区域免受不必要的干扰的需求变得更加重要。在这项研究中，我们采用了双报告工具与 CyanoGate MoClo Assembly 系统配合使用，该系统可以量化和比较不同物种内部和之间终止子序列的效率。我们鉴定了大肠杆菌、集胞藻属中的 34 个内在终止子。 PCC 6803 和细长聚球藻 UTEX 2973 并观察到终止效率的显着差异。然而，我们还在所有三个物种中鉴定了五个终止效率> 96％的终止子，这表明一些终止子在异养物种和蓝细菌中都可以表现一致。
Cyanobacteria utilize sunlight to convert carbon dioxide into a wide variety of secondary metabolites and show great potential for green biotechnology applications. Although cyanobacterial synthetic biology is less mature than for other heterotrophic model organisms, there are now a range of molecular tools available to modulate and control gene expression. One area of gene regulation that still lags behind other model organisms is the modulation of gene transcription, particularly transcription termination. A vast number of intrinsic transcription terminators are now available in heterotrophs, but only a small number have been investigated in cyanobacteria. As artificial gene expression systems become larger and more complex, with short stretches of DNA harboring strong promoters and multiple gene expression cassettes, the need to stop transcription efficiently and insulate downstream regions from unwanted interference is becoming more important. In this study, we adapted a dual reporter tool for use with the CyanoGate MoClo Assembly system that can quantify and compare the efficiency of terminator sequences within and between different species. We characterized 34 intrinsic terminators in Escherichia coli, Synechocystis sp. PCC 6803, and Synechococcus elongatus UTEX 2973 and observed significant differences in termination efficiencies. However, we also identified five terminators with termination efficiencies of >96% in all three species, indicating that some terminators can behave consistently in both heterotrophic species and cyanobacteria.
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影响因子：
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作者：
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影响因子：
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作者：
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