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硅藻中在低盐度条件下活跃的VOC启动子的表征

Characterization of the VOC Promoter That Is Active Under Low-Salinity Conditions in the Diatom .

作者信息

Toustou Charlotte, Plasson Carole, Kiefer-Meyer Marie-Christine, Bardor Muriel

机构信息

Laboratoire Glycobiologie et Matrice Extracellulaire Végétale (GlycoMEV) UR4358, University of Rouen Normandie (UNIROUEN), Normandie Université, 76821 Mont-Saint-Aignan, France.

ALGA BIOLOGICS, Centre Universitaire de Recherche et d'Innovation en Biologie (CURIB), 25 Rue Tesnière, 76821 Mont-Saint-Aignan, France.

出版信息

Mar Drugs. 2025 Apr 26;23(5):185. doi: 10.3390/md23050185.

Abstract

Microalgae such as are promising cell biofactories for the production of high-value molecules, including monoclonal antibodies (mAbs). However, to date, the production of mAbs in using the inducible nitrate reductase (NR) promoter has yielded only a limited amount of mAbs. Therefore, the identification of a robust promoter that produces high yields of mAbs is crucial for the development of a cost-effective expression system. To date, only a few endogenous promoters have been characterized in . In this study, we identified thirty-three potential "strong" endogenous promoters based on our previously published transcriptomic data from the Pt3 strain. These putative promoter sequences were cloned into an episomal vector and fused to the gene encoding enhanced green fluorescent protein (eGFP). Their strength was assessed by measuring eGFP fluorescence, which reflects the level of eGFP protein expression. Of the thirty-three promoters, thirteen were able to successfully drive eGFP protein expression. Among them, the best results were obtained with the VOC promoter, which allowed a significant increase in eGFP expression compared to that induced by the NR promoter. These results contribute to the identification of new genetic tools that can be used in future studies to increase the yield of production of recombinant proteins in at an industrial scale.

摘要

诸如 之类的微藻是生产包括单克隆抗体(mAbs)在内的高价值分子的有前景的细胞工厂。然而,迄今为止,使用诱导型硝酸还原酶(NR)启动子在 中生产单克隆抗体的产量仍然有限。因此,鉴定出能高产单克隆抗体的强大启动子对于开发具有成本效益的表达系统至关重要。迄今为止,在 中仅鉴定了少数内源性启动子。在本研究中,我们基于之前发表的来自Pt3菌株的转录组数据,鉴定了33个潜在的“强”内源性启动子。这些假定的启动子序列被克隆到一个附加型载体中,并与编码增强型绿色荧光蛋白(eGFP)的基因融合。通过测量反映eGFP蛋白表达水平的eGFP荧光来评估它们的强度。在这33个启动子中,有13个能够成功驱动eGFP蛋白表达。其中,VOC启动子取得了最佳结果,与NR启动子诱导的表达相比,它能使eGFP表达显著增加。这些结果有助于鉴定新的遗传工具,可用于未来的研究中,以在工业规模上提高 在重组蛋白生产中的产量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e16c/12113381/02a49167982c/marinedrugs-23-00185-g001.jpg

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