Zymomonas mobilis is a model bacterial ethanologen and has been engineered to produce lignocellulosic biofuels and biochemicals such as 2,3-butanediol. We have previously identified promoters of different strengths using systems biology datasets and characterized them using the flow cytometry-based dual reporter-gene system. Here, we further demonstrated the capability of applying the dual reporter-gene system and omics datasets on discovering inducible promoters. Ten candidate ethanol-inducible promoters were identified through omics datasets mining and clustering. Using the dual reporter-gene system, these promoters were characterized under natural growth, ethanol stress, and ethanol-induced condition to investigate the transcriptional strength and ethanol inducibility. The results demonstrated that three promoters of P0405, P0435, and P0038 driving the expression of native genes of ZMO0405, ZMO0435, and ZMO0038, correspondingly, are potential ethanol-responsive promoters and may be growth related. This study not only identified and verified three ethanol-inducible promoters as biological parts, which can be used to synchronize the expression of heterologous pathway genes with the ethanol production process of Z. mobilis, but also demonstrated the power of combining omics datasets and dual reporter-gene system to identify biological parts for metabolic engineering and synthetic biology applications in Z. mobilis and related microorganisms.

中文翻译：

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基于组学数据和双报告基因系统，鉴定和鉴定运动发酵单胞菌乙醇诱导型启动子。

运动发酵单胞菌（Zymomonas mobilis）是一种典型的细菌乙醇原，已被工程化以生产木质纤维素生物燃料和生物化学物质，例如2,3-丁二醇。我们先前使用系统生物学数据集确定了不同强度的启动子，并使用基于流式细胞仪的双重报告基因系统对其进行了表征。在这里，我们进一步证明了在发现诱导型启动子上应用双重报告基因系统和组学数据集的能力。通过组学数据集挖掘和聚类，确定了十种候选乙醇诱导型启动子。使用双报告基因系统，这些启动子在自然生长，乙醇胁迫和乙醇诱导的条件下进行了表征，以研究转录强度和乙醇诱导性。结果表明，P0405，P0435，驱动ZMO0405，ZMO0435和ZMO0038的天然基因表达的P1038和P0038分别是潜在的乙醇应答性启动子，并且可能与生长有关。这项研究不仅鉴定并验证了三种乙醇诱导型启动子作为生物部分，可用于使异源途径基因的表达与运动发酵单胞菌的乙醇生产过程同步，而且证明了结合组学数据集和双重报告基因的功能。 -基因系统，用于识别运动发酵单胞菌及相关微生物中代谢工程和合成生物学应用的生物部分。