Weichert Nicola, Hauptmann Valeska, Helmold Christine, Conrad Udo
Department of Molecular Genetics, Leibniz Institute of Plant Genetics and Crop Plant Research Gatersleben, Germany.
Front Plant Sci. 2016 Jan 28;7:6. doi: 10.3389/fpls.2016.00006. eCollection 2016.
Seeds enable plants to germinate and to grow in situations of limited availability of nutrients. The stable storage of different seed proteins is a remarkable presumption for successful germination and growth. These strategies have been adapted and used in several molecular farming projects. In this study, we explore the benefits of seed-based expression to produce the high molecular weight spider silk protein FLAG using intein-based trans-splicing. Multimers larger than 460 kDa in size are routinely produced, which is above the native size of the FLAG protein. The storage of seeds for 8 weeks and 1 year at an ambient temperature of 15°C does not influence the accumulation level. Even the extended storage time does not influence the typical pattern of multimerized bands. These results show that seeds are the method of choice for stable accumulation of products of complex transgenes and have the capability for long-term storage at moderate conditions, an important feature for the development of suitable downstream processes.
种子使植物能够在营养物质供应有限的情况下发芽和生长。不同种子蛋白的稳定储存是成功发芽和生长的显著前提。这些策略已在多个分子农业项目中得到应用。在本研究中，我们探索了基于种子表达利用内含肽介导的反式剪接来生产高分子量蜘蛛丝蛋白FLAG的益处。通常会产生大小超过460 kDa的多聚体，这高于FLAG蛋白的天然大小。在15°C的环境温度下将种子储存8周和1年不会影响积累水平。即使延长储存时间也不会影响多聚化条带的典型模式。这些结果表明，种子是稳定积累复杂转基因产物的首选方法，并且具有在适度条件下长期储存的能力，这是开发合适下游工艺的一个重要特征。
