Sánchez Flora, Ponz Fernando
Centro de Biotecnología y Genómica de Plantas (CBGP, UPM-INIA), Madrid, Spain.
Methods Mol Biol. 2018;1776:471-485. doi: 10.1007/978-1-4939-7808-3_31.
Potyviruses are plant viruses with elongated, flexuous virions amenable to modifications in the only viral structural protein, the coat protein (CP). Out of the several theoretically possible modifications to the CP, the one most exploited for peptide presentation is the genetic fusion of the peptide-to-be-expressed, to the CP N-terminus. Successful high-level expression of the modified CP has been achieved this way. The purified recombinant viral particles incorporate most, if not all, the properties of the expressed peptides. For many purposes, the recombinant virus particles present in extracts of infected plants should be purified for further use. Procedures for carrying out the whole process, from cloning to purification are described in the chapter.
马铃薯Y病毒属病毒是一类植物病毒，其病毒粒子呈细长、弯曲状，易于在唯一的病毒结构蛋白即外壳蛋白（CP）上进行修饰。在对CP的几种理论上可行的修饰中，用于肽展示的最常用方法是将待表达的肽与CP的N端进行基因融合。通过这种方式已成功实现了修饰后CP的高水平表达。纯化的重组病毒粒子具有所表达肽的大部分（如果不是全部）特性。出于多种目的，感染植物提取物中存在的重组病毒粒子应进行纯化以供进一步使用。本章描述了从克隆到纯化的整个过程的操作步骤。
