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Journal of Integrative Agriculture  2019, Vol. 18 Issue (10): 2361-2368    DOI: 10.1016/S2095-3119(19)62717-6
Animal Science · Veterinary Medicine Advanced Online Publication | Current Issue | Archive | Adv Search |
Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection
ZHAO Dan-dan1*, SHUAI Lei1*, GE Jin-ying1, WANG Jin-liang1, WEN Zhi-yuan1, LIU Ren-qiang1, WANG Chong1, WANG Xi-jun1, BU Zhi-gao1, 2      
1 State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150069, P.R.China
2 Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, P.R.China
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The mechanism of rabies virus (RABV) infection still needs to be further characterized.  RABV particle with self-fluorescent is a powerful viral model to visualize the viral infection process in cells.  Herein, based on a reverse genetic system of the Evelyn-Rokitnicki-Abelseth (rERA) strain, we generated a recombinant RABV rERA-N/mCherry strain that stably expresses an additional ERA nucleoprotein that fuses with the red fluorescent protein mCherry (N/mCherry).  The rERA-N/mCherry strain retained growth property similar to the parent strain rERA in vitro.  The N/mCherry expression showed genetic stability during passage into mouse neuroblastoma (NA) cells and did not change the virulence of the vector.  The rERA-N/mCherry strain was then utilized as a visual viral model to study the RABV-cell binding and internalization.  We directly observed the red self-fluorescence of rERA-N/mCherry particles binding to the cell surface, and further co-localizing with clathrin in the early stage of infection in NA cells by fluorescence microscopy.  Our results showed that the rERA-N/mCherry strain uses clathrin-dependent endocytosis to enter cells, which is consistent with the well-known mechanism of RABV invasion.  The recombinant RABV rERA-N/mCherry thus appears to have the potential to be an effective viral model to further explore the fundamental molecular mechanism of rabies neuropathogenesis.
Keywords:  rabies virus        self-fluorescence       binding        internalization  
Received: 14 January 2019   Accepted: 29 September 2019
Fund: This work was supported by the National Natural Science Fundation of China (31800138) and the National Key Research and Development Program of China (2016YFD0500403).
Corresponding Authors:  Correspondence BU Zhi-gao, E-mail:; WANG Xi-jun, E-mail:    
About author:  ZHAO Dan-dan, E-mail:; SHUAI Lei, E-mail:; * These authors contributed equally to this study.

Cite this article: 

ZHAO Dan-dan, SHUAI Lei, GE Jin-ying, WANG Jin-liang, WEN Zhi-yuan, LIU Ren-qiang, WANG Chong, WANG Xi-jun, BU Zhi-gao. 2019. Generation of recombinant rabies virus ERA strain applied to virus tracking in cell infection. Journal of Integrative Agriculture, 18(10): 2361-2368.

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