植物瞬时表达生产和纯化病毒样颗粒。
Production and Purification of Virus-Like Particles by Transient Expression in Plants.
机构信息
Centre for Cell Factories and Biopolymers, Griffith Institute for Drug Discovery, Griffith University, Nathan, QLD, Australia.
出版信息
Methods Mol Biol. 2023;2671:387-402. doi: 10.1007/978-1-0716-3222-2_22.
Transient expression in plants has become a useful production system for virus-like particle (VLP) expression. High yields and flexible approaches to assembling complex VLPs, combine with ease of scale-up and inexpensive reagents to provide an attractive method for recombinant protein expression in general. Plants have demonstrated excellent capacity for the assembly and production of protein cages for use in vaccine design and nanotechnology. Furthermore, numerous virus structures have now been determined using plant-expressed VLPs, showing the utility of this approach in structural virology. Transient protein expression in plants uses common microbiology techniques, leading to a straightforward transformation procedure that does not result in stable transgenesis. In this chapter, we aim to provide a generic protocol for transient expression of VLPs in Nicotiana benthamiana using soil-free plant cultivation and a simple vacuum infiltration procedure, along with methodology for purifying VLPs from plant leaves.
在植物中进行瞬时表达已成为病毒样颗粒 (VLP) 表达的一种有用的生产系统。高产量和灵活的方法来组装复杂的 VLPs,结合易于扩大规模和廉价的试剂,为一般的重组蛋白表达提供了一种有吸引力的方法。植物已经展示了在疫苗设计和纳米技术中组装和生产蛋白质笼的出色能力。此外,现在已经使用植物表达的 VLPs 确定了许多病毒结构,表明了这种方法在结构病毒学中的实用性。植物中的瞬时蛋白质表达使用常见的微生物学技术,导致直接的转化程序,而不会导致稳定的转基因。在本章中,我们旨在提供一种使用无土植物培养和简单的真空渗透程序在 Nicotiana benthamiana 中瞬时表达 VLPs 的通用方案,以及从植物叶片中纯化 VLPs 的方法。