Purification of His-Tagged Proteases from the Apoplast of Agroinfiltrated N. benthamiana.
Purification of His-Tagged Proteases from the Apoplast of Agroinfiltrated N. benthamiana.
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从农杆菌渗透的本塞姆氏烟草的质外体中纯化组氨酸标记的蛋白酶。
DOI：
10.1007/978-1-0716-2079-3_5
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Schuster M
中科院分区：
文献类型：
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作者：
Schuster M
通过农杆菌渗入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质的生化表征。在这一章中，我们描述了纯化分泌的，组氨酸标记的蛋白酶从质外体agroinfiltratedNicotiana benthamianausing固定化金属亲和层析（IMAC）。我们展示了纯化蛋白酶的质量检查，并讨论了潜在的问题和规避它们的方法。作为概念验证，我们生产和纯化番茄免疫蛋白酶Pip1，并证明纯化后的蛋白质是有活性的。
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 agroinfiltratedNicotiana benthamianausing 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.