从农杆菌浸润的 N. benthamiana 质外体中纯化 His 标记的蛋白酶。
Purification of His-Tagged Proteases from the Apoplast of Agroinfiltrated N. benthamiana.
机构信息
The Plant Chemetics Laboratory, Department of Plant Sciences, University of Oxford, Oxford, UK.
出版信息
Methods Mol Biol. 2022;2447:53-66. doi: 10.1007/978-1-0716-2079-3_5.
Protein expression in plants by agroinfiltration and subsequent purification is increasingly used for the biochemical characterization of plant proteins. In this chapter we describe the purification of secreted, His-tagged proteases from the apoplast of agroinfiltrated Nicotiana benthamiana using immobilized metal affinity chromatography (IMAC). We show quality checks for the purified protease and discuss potential problems and ways to circumvent them. As a proof of concept, we produce and purify tomato immune protease Pip1 and demonstrate that the protein is active after purification.
农杆菌浸润和随后的纯化在植物中表达蛋白质，越来越多地用于植物蛋白的生化特性分析。在这一章中，我们描述了使用固定化金属亲和层析（IMAC）从农杆菌浸润的烟草细胞外区域中纯化分泌的、His 标签的蛋白酶。我们展示了对纯化蛋白酶的质量检查，并讨论了潜在的问题和解决方法。作为一个概念验证，我们生产和纯化了番茄免疫蛋白酶 Pip1，并证明了该蛋白在纯化后具有活性。
Zotero 插件