单病毒示踪技术在动植物细胞中的应用进展

doi:10.13560/j.cnki.biotech.bull.1985.2023-1044

摘要/Abstract

摘要：

病毒（virus）是一种专营胞内寄生的简单微生物，是造成动植物病害的主要病原体之一。病毒的侵染过程高度动态且复杂，单病毒示踪技术的发展为揭示病毒生命活动的精细过程提供了可能。单病毒示踪是一种基于荧光标记与高时空分辨率成像的新方法，允许对单一或多个病毒的生命活动进行观察。近几年，单病毒示踪技术在病毒入胞、复制、细胞间传染等方面的研究中获得广泛应用，使病毒感染机制的研究取得较大的进展。本文介绍了病毒入侵细胞机制的研究进展, 重点总结了荧光标记在单病毒示踪上的应用, 特别是对病毒不同结构的标记和成像方法进行了详细论述, 以期为优化标记策略、解决对不同种类和结构的病毒实现高效标记等问题提供新思路。

关键词: 病毒标记, 单病毒示踪, 荧光染料, 荧光蛋白, 量子点

Abstract:

Virus is a kind of simple microorganism specializing in intracellular parasitics, which is one of the main pathogens causing animal and plant diseases. The development of single-virus tracking（SVT）technology makes it possible to reveal the fine process of viral life activity. SVT is a new method to study the mechanism of viral infection based on fluorescence labeling and high spatio-temporal resolution imaging, which allows the observation of the life activity of a single or multiple viruses. In recent years, the SVT technology has made great progress in the study of virus entry, replication, and intercellular transmission, providing new insights into the mechanism of virus infection. In this paper, the principle and strategy types of virus labeling are briefly introduced, and the research progress of virus fluorescence labeling in single virus tracing is summarized, especially the labeling and imaging methods of different structures of viruses, aiming to provide new ideas for optimizing labeling strategies and solving the problem of efficient labeling of different types and structures of viruses.

Key words: virus labeling, single virus tracking, fluorescent dye, fluorescent protein, quantum dots

袁笑妍, 张御格, 姚丽娟, 唐晨, 李晓娟. 单病毒示踪技术在动植物细胞中的应用进展[J]. 生物技术通报, 2024, 40(4): 52-66.

YUAN Xiao-yan, ZHANG Yu-ge, YAO Li-juan, TANG Chen, LI Xiao-juan. Application of Single-virus Tracking Technique in Animal and Plant Cells[J]. Biotechnology Bulletin, 2024, 40(4): 52-66.

图/表 3

图1 动物病毒和植物病毒的生命周期动物病毒经过受体接触、入胞、转运、脱壳、复制、翻译、组装等一系列步骤合成子代病毒，通过胞吐、出芽或裂解细胞的方式出胞；植物病毒在入侵细胞后经历和动物病毒相似的过程，不同的是在完成子代病毒组装后，经由植物的胞间连丝出胞，进行细胞间运输

Fig. 1 Life cycles of animal virus and plant virus After a series of steps such as receptor contact, entry, transport, uncoating, replication, translation and assembly, the animal virus synthesized the progeny virus and left the cell through exocytosis, budding or lysis. Plant viruses undergo a similar process to animal viruses after entering the cells. The difference is that they leave the primary infected cells through the plant plasmodesmata and carry out intercellular transport after completing the assembly of progeny viruses

图2 病毒荧光标记策略单病毒示踪技术中常用的病毒荧光标记的策略主要可以分为化学法、标签法和编码荧光蛋白法三类。化学法包括通过交联剂偶联法、点击化学法和生物素-链霉亲和素互作法三种；标签法包括肽标签和核酸标签两种

Fig. 2 Virus fluorescence labeling strategies The commonly used fluorescence labeling strategies in single virus tracking technology can be divided into three categories: Chemical labeling, coding tag labeling and encoding fluorescent protein labeling. Chemical labeling methods include cross-linking reaction, click chemistry and biotin-streptavidin interaction; coding tag labeling methods include peptide tag-mediated labeling and oligonucleotide-guided labeling

图3 不同成像技术的时空分辨率从上至下依次展示了单病毒示踪实验中常用荧光显微成像技术时空分辨率范围。修改自Schermelleh 等[125]

Fig. 3 Spatiotemporal resolution of different imaging techniques The figure shows, from top to bottom, the range of spatial and temporal resolution of the fluorescent microscopy imaging techniques commonly used in single-virus tracking. Refer to Schermelleh et al[125]

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