[1]单博文,王金良,刘仁强,等.红色荧光示踪狂犬病病毒ERA株病毒粒子方法的建立[J].中国预防兽医学报,2018,(02):112-116.[doi:0.3969/j.issn.1008-0589.201704028]
 SHAN Bo-wen,WANG Jin-liang,LIU Ren-qiang,et al.Conjugation of Rabies virus ERA strain particle with CY5-NHS-ester for virus tracking in cell invasion[J].Chinese journal of preventive veterinary medicine,2018,(02):112-116.[doi:0.3969/j.issn.1008-0589.201704028]
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红色荧光示踪狂犬病病毒ERA株病毒粒子方法的建立()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2018年02
页码:
112-116
栏目:
诊断和检测技术
出版日期:
2018-02-15

文章信息/Info

Title:
Conjugation of Rabies virus ERA strain particle with CY5-NHS-ester for virus tracking in cell invasion
文章编号:
1008-0589(2018)2-0112-05
作者:
 

单博文王金良刘仁强帅 磊王喜军葛金英温志远夏咸柱步志高*

 (中国农业科学院哈尔滨兽医研究所 兽医生物技术国家重点实验室,黑龙江 哈尔滨 150069)
Author(s):
 

SHAN Bo-wen WANG Jin-liang LIU Ren-qiang SHUAI Lei WANG Xi-jun GE Jin-yingWEN Zhi-yuan XIA Xian-zhu BU Zhi-gao*

 (State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences,
Harbin 150069, China)
关键词:
狂犬病病毒ERA株标记活细胞成像
Keywords:
 rabies virus  ERA strain  staining  live cell imaging
分类号:
S852.65
DOI:
0.3969/j.issn.1008-0589.201704028
文献标志码:
A
摘要:
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。
  
Abstract:
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 viruses.

参考文献/References:

[1] Dietzschold B, Schnell M, Koprowski H. Pathogenesis of Rabies[J]. CTMI, 2005, 292: 45-56.
[2]Faber M, Li J, Kean R B, et al. Effective preexposure and postexposure prophylaxis of rabies with a highly attenuated recombinant rabies virus [J]. PNAS, 2009, 106(27): 11300-11305.
[3]Dietzschold B, Wunner W H, Wiktor T J, et al. Characterization of an antigenic determinant of the glycoprotein that correlates with pathogenicity of rabies virus [J]. PNAS, 1983, 80(1): 70-74.
[4]Klingen Y, Conzelmann K K, Finke S. Double-labeled rabies virus: live tracking of enveloped virus transport [J]. J Virol, 2008, 82(1): 237-245.
[5]Schaar H M V D, Rust M J, Chen C, et al. Dissecting the cell entry pathway of dengue virus by single-particle tracking in living cells [J]. PLoS Pathog, 2008, 4(12): e1000244.
[6]Lakadamyali M, Rust M J, Babcock H P, et al. Visualizing infection of individual influenza viruses [J]. PNAS, 2003, 100(16): 9280-9285.
[7]Mcdonald D, Vodicka M A, Lucero G, et al. Visualization of the intracellular behavior of HIV in living cells [J]. J Cell Biol, 2002, 159(3): 441.
[8]Brandenburg B, Lee L Y, Lakadamyali M, et al. Imaging poliovirus entry in live cells [J]. PLoS Biol, 2007, 5(7): e183.
[9]Zhou Peng, Zheng Zhen-hua, Lu Wen, et al. Multicolor labeling of living virus particles in live cells [J]. Angew Chem Int Ed Engl, 2012, 51(3): 670-674.
[10]Xu Hai-jiao, Hao Xian, Wang Shao-wen, et al. Real-time imaging of rabies virus entry into living vero cells [J]. Sci Rep, 2015, 5: 11753.
[11]周微微,王金玲,祖述龙,等. 2009年~2012年内蒙古地区狂犬病毒分离株遗传变异分析与免疫保护效果研究[J]. 中国预防兽医学报,2016,38(6):458-462.
                             (本文编辑:彭永刚)

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更新日期/Last Update: 2018-03-09