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基于理性设计的 中的蓝光响应性强合成启动子

A Blue Light-Responsive Strong Synthetic Promoter Based on Rational Design in .

机构信息

Guangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen Engineering Laboratory for Marine Algal Biotechnology, Longhua Innovation Institute for Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China.

出版信息

Int J Mol Sci. 2023 Sep 27;24(19):14596. doi: 10.3390/ijms241914596.

Abstract

() is a single-cell green alga that can be easily genetically manipulated. With its favorable characteristics of rapid growth, low cost, non-toxicity, and the ability for post-translational protein modification, has emerged as an attractive option for the biosynthesis of various valuable products. To enhance the expression level of exogenous genes and overcome the silencing of foreign genes by , synthetic promoters such as the chimeric promoter AR have been constructed and evaluated. In this study, a synthetic promoter GA was constructed by hybridizing core fragments from the natural promoters of the acyl carrier protein gene () and the glutamate dehydrogenase gene (). The GA promoter exhibited a significant increase (7 times) in expressing GUS, over the AR promoter as positive control. The GA promoter also displayed a strong responsiveness to blue light (BL), where the GUS expression was doubled compared to the white light (WL) condition. The ability of the GA promoter was further tested in the expression of another exogenous cadA gene, responsible for catalyzing the decarboxylation of lysine to produce cadaverine. The cadaverine yield driven by the GA promoter was increased by 1-2 times under WL and 2-3 times under BL as compared to the AR promoter. This study obtained, for the first time, a blue light-responsive GDH2 minimal fragment in , which delivered a doubling effect under BL when used alone or in hybrid. Together with the strong GA synthetic promoter, this study offered useful tools of synthetic biology to the algal biotechnology field.

摘要

()是一种单细胞绿藻,易于进行基因操作。由于其生长迅速、成本低、无毒、可进行翻译后蛋白修饰等特点,已成为生物合成各种有价值产物的有吸引力的选择。为了提高外源基因的表达水平并克服基因沉默,已经构建和评估了合成启动子,如嵌合启动子 AR。在这项研究中,通过杂交酰基载体蛋白基因()和谷氨酸脱氢酶基因()的天然启动子的核心片段,构建了一个合成启动子 GA。与阳性对照的 AR 启动子相比,GA 启动子显著提高了 GUS 的表达(7 倍)。GA 启动子对蓝光(BL)也表现出强烈的响应性,与白光(WL)条件相比,GUS 表达增加了一倍。进一步在表达另一个负责催化赖氨酸脱羧产生尸胺的外源 cadA 基因时测试了 GA 启动子的能力。与 AR 启动子相比,在 WL 和 BL 下,GA 启动子驱动的尸胺产量分别增加了 1-2 倍和 2-3 倍。本研究首次在中获得了一个蓝光响应的 GDH2 最小片段,单独或杂交使用时在 BL 下具有倍增效果。与强大的 GA 合成启动子一起,这项研究为藻类生物技术领域提供了有用的合成生物学工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa8f/10572394/182e5add32c2/ijms-24-14596-g001.jpg

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