红色荧光示踪狂犬病病毒ERA株病毒粒子方法的建立

摘要

To establish a method for staining rabies virus to observe how the ERA strain of rabies virus (RABV) infect living cells and shelling,the amino-group of RABV was stained into red by CY5-NHS-ester,the cell membrane was dyed into green by lipophilic carbonyl cyanine fluorescent dyes and the cell nuclei was dyed into bright blue by DNA fluorescent dyes.The adsorption and endocytosis process of the ERA strain of rabies virus could be clearly observed by using 3D Live Cell Imaging System.The Cy5 staining solution suits for staining rabies virus to observe and screen the viruses when they are infecting cells with excellent staining results.This experiment provids an effective way to study the pathogenesis of RABV and also lay a foundation to establishment of staining methods for other Rhaboviridae vimses.%为建立一种狂犬病病毒(RABV)荧光标记方法,便于有效地观察研究RABV侵入细胞及脱壳的过程,本研究采用CY5-NHS酯红色荧光染料标记氨基的方法将纯化的RABV ERA株进行CY5-NHS酯荧光染料标记;采用亲脂性羰花青荧光染料将细胞膜染成绿色;采用DNA荧光染料将细胞核染成亮蓝色;在3D活细胞激光共聚焦分析成像系统下成像,动态观察病毒吸附细胞并内吞的过程.与传统的染色方法相比,本研究建立的病毒标记方法更适合在3D活细胞激光共聚焦分析成像系统下观察和拍摄RABV感染细胞的过程,为RABV感染机制的研究提供了一种有效研究方法,对于其它弹状病毒科病毒标记方法的建立也具有一定的参考意义.