Rapid Generation of Universal Synthetic Promoters for Controlled Gene Expression in Both Gas-Fermenting and Saccharolytic Clostridium Species.
Rapid Generation of Universal Synthetic Promoters for Controlled Gene Expression in Both Gas-Fermenting and Saccharolytic Clostridium Species.
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DOI：
10.1021/acssynbio.7b00155
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发表时间：
2017-06
影响因子：
4.7
通讯作者：
Gaohua Yang;Dechen Jia;Lin Jin;Yuqian Jiang;Yong Wang;Weihong Jiang;Yang Gu
中科院分区：
文献类型：
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作者：
Gaohua Yang;Dechen Jia;Lin Jin;Yuqian Jiang;Yong Wang;Weihong Jiang;Yang Gu
工程溶剂型梭状芽孢杆菌是一类重要的工业微生物，由于缺乏丰富的生物成分，其在生物精炼中的应用潜力难以充分发挥。在这里，我们开发了一种基于双报告系统(CatP-LacZ)和广泛使用的强Thl启动子的快速构建溶血性梭状芽孢杆菌合成启动子文库的有效方法。得到的人工启动子跨度为2个数量级，由两个模块组成(核心启动子区域和RBS与翻译起始码之间的间隔区)，最强的启动子的活性是原始表达部分Pth1的10倍以上。将这些合成的启动子分别在糖解菌和气体发酵菌中控制表达SADH和Dank，得到了预期的表型，并且启动子活性与表型变化有很好的相关性。本文提出的大强度范围启动子将有助于合成生物学在溶血性梭状芽孢杆菌中的应用。
Engineering solventogenic clostridia, a group of important industrial microorganisms, to realize their full potential in biorefinery application is still hindered by the absence of plentiful biological parts. Here, we developed an effective approach for rapid generation of a synthetic promoter library in solventogenic clostridia based on a dual-reporter system (catP-lacZ) and a widely used strong thl promoter. The yielded artificial promoters, spanning 2 orders of magnitude, comprised two modular components (the core promoter region and the spacer between RBS and the translation-initiating code), and the strongest promoter had an over 10-fold-higher activity than the original expression part Pthl. The test of these synthetic promoters in controlled expression of sadh and danK in saccharolytic C. acetobutylicum and gas-fermenting C. ljungdahlii, respectively, gave the expected phenotypes, and moreover, showed good correlation between promoter activities and phenotypic changes. The presented wide-strength-range promoters here will be useful for synthetic biology application in solventogenic clostridia.