Design and Reconstruction of Regulatory Parts for Fast-frowing Vibrio natriegens Synthetic Biology.,ACS Synthetic Biology

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Design and Reconstruction of Regulatory Parts for Fast-frowing Vibrio natriegens Synthetic Biology.
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-07-28 , DOI: 10.1021/acssynbio.0c00158
Fengli Wu _{1,

2} , Wujiu Chen ₁ , Yanfeng Peng ₁ , Ran Tu ₁ , Yuping Lin ₁ , Jianmin Xing ₂ , Qinhong Wang ₁

Affiliation

The fast-growing Vibrio natriegens is an attractive robust chassis for diverse synthetic biology applications. However, V. natriegens lacks the suitable constitutive regulatory parts for precisely tuning the gene expression and, thus, recapitulating physiologically relevant changes in gene expression levels. In this study, we designed, constructed, and screened the synthetic regulatory parts by varying the promoter region and ribosome binding site element for V. natriegens with different transcriptional or translational strengths, respectively. The fluorescence intensities of the cells with different synthetic regulatory parts could distribute evenly over a wide range of 5 orders of magnitude. The selected synthetic regulatory parts had good stability in both nutrient-rich and minimal media. The precise combinatorial modulation of galP (GalP = galactose permease) and glk (Glk = glucokinase) from Escherichia coli by using three synthetic regulatory parts with different strengths was confirmed in a phosphoenolpyruvate:carbohydrate phosphotransferase system with inactive V. natriegens strain to alter the glucose transport. This work provides the simple, efficient, and standardized constitutive regulatory parts for V. natriegens synthetic biology.

中文翻译：

速生弧菌合成生物学调控部件的设计与重构。

快速增长的Vibrio natriegens是适用于各种合成生物学应用的有吸引力的坚固底盘。然而，V. natriegens缺乏合适的组成性调控部分来精确调整基因表达，从而重现基因表达水平的生理相关变化。在这项研究中，我们通过改变V. natriegens的启动子区域和核糖体结合位点元件来设计、构建和筛选合成调控部分分别具有不同的转录或翻译强度。具有不同合成调节部分的细胞的荧光强度可以在 5 个数量级的广泛范围内均匀分布。选定的合成调节部分在营养丰富的培养基和基本培养基中均具有良好的稳定性。在磷酸烯醇式丙酮酸：含无活性钠酸弧菌的碳水化合物磷酸转移酶系统中，证实了通过使用三种不同强度的合成调控部分对来自大肠杆菌的galP（GalP = 半乳糖通透酶）和glk（Glk = 葡糖激酶）的精确组合调节应变以改变葡萄糖转运。这项工作为V. natriegens合成生物学提供了简单、高效和标准化的组成调节部分。

更新日期：2020-09-20

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