利用进化的 RNA 聚合酶拆分体进行农业化学控制基因表达。II。
Agrochemical control of gene expression using evolved split RNA polymerase. II.
机构信息
Department of Neurophysiology and Neuropharmacology, Institute of Special Environmental Medicine and Co-innovation Center of Neuroregeneration, Nantong University, Nantong, China.
Duke Kunshan University, Kunshan, Jiangsu Province, China.
出版信息
PeerJ. 2024 Sep 4;12:e18042. doi: 10.7717/peerj.18042. eCollection 2024.
Agrochemical inducible gene expression system provides cost-effective and orthogonal control of energy and information flow in bacterial cells. However, the previous version of Mandipropamid inducible gene expression system (Mandi-T7) became constitutively active at room temperature. We moved the split site of the eRNAP from position LYS179 to position ILE109. This new eRNAP showed proximity dependence at 23 °C, but not at 37 °C. We built Mandi-T7-v2 system based on the new eRNAP and it worked in both and . We also induced GFP expression in cells in a semi- system. The modified eRNAP when combined with the leucine zipper-based dimerization system, behaved as a cold inducible gene expression system. Our new system provides a means to broaden the application of agrochemicals for both research and agricultural application. Portions of this text were previously published as part of a preprint (https://www.biorxiv.org/content/10.1101/2024.04.02.587689v1).
农用化学品诱导基因表达系统为控制细菌细胞的能量和信息流提供了具有成本效益和正交的控制手段。然而，以前的 Mandipropamid 诱导基因表达系统（Mandi-T7）在室温下变为组成型激活。我们将 eRNAP 的分裂位点从位置 LYS179 移动到位置 ILE109。这种新的 eRNAP 在 23°C 时表现出接近依赖性，但在 37°C 时没有。我们基于新的 eRNAP 构建了 Mandi-T7-v2 系统，它在 和 中都能正常工作。我们还在 细胞中诱导 GFP 表达，建立了一个半诱导系统。当与基于亮氨酸拉链的二聚化系统结合时，经过修饰的 eRNAP 表现为冷诱导基因表达系统。我们的新系统为农用化学品的研究和农业应用提供了一种拓宽应用的手段。本文的部分内容之前已作为预印本的一部分发表（https://www.biorxiv.org/content/10.1101/2024.04.02.587689v1）。
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