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| 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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Abstract 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.
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Received: 14 January 2019
Accepted: 29 September 2019
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| 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). |
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Corresponding Authors:
Correspondence BU Zhi-gao, E-mail: buzhigao@caas.cn; WANG Xi-jun, E-mail: wangxijun@caas.cn
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| About author: ZHAO Dan-dan, E-mail: 13633636807@163.com; SHUAI Lei, E-mail: shuailei@caas.cn; * 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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