Effect of D393E mutation in envelope protein of Japanese encephalitis/Zika chimeric virus on the intracerebral neurovirulence in mice
10.3760/cma.j.cn112866-20200417-00133
- VernacularTitle:寨卡病毒包膜蛋白D393E突变对乙脑/寨卡嵌合病毒小鼠脑内神经毒力的影响
- Author:
Yalan FENG
1
;
Yueke LI
;
Yang REN
;
Rong HUANG
;
Lei YUAN
;
Liping TANG
;
Jian YANG
Author Information
1. 川北医学院基础医学院，南充 637000
- Keywords:
Zika virus;
Chimeric virus;
Neurovirulence
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(4):391-395
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explore the effect of D393E mutation on its intracerebral neurovirulence in mice after infectious clone of the chimeric Japanese encephalitis virus (JEV)/Zika virus (ZIKV) containing envelope protein D393E mutant was built and the virus was rescued.Methods:The full-length cDNA plasmid of JEV/ZIKV chimeric virus containing D393E mutation in the envelope protein was built by SOE PCR and gene cloning technology. The linearized fragment of the plasmid was used for in vitro transcription to obtain viral RNA, followed by electrotransfection into BHK-21 cells to rescue the mutant virus JEV/ZIKV(D393E). The plaque size and growth characteristics of JEV/ZIKV (D393E) and JEV/ZIKV were compared after BHK-21 cells were inoculated with the viruses. The intracerebral neurovirulence of the viruses was calculated after the viruses were inoculated in mice intracerebrally (i.c.).Results:Restriction enzyme digestion evaluation confirmed that the infectious clone of mutant virus was correctly built. The plaque assay demonstrated that the plaque diameter of JEV/ZIKV (D393E) was about 0.7±0.2 mm, which was smaller than that of JEV/ZIKV (1.3±0.2 mm). The LD 50 value of the mutant was 31.51 PFU when injected in mice via i. c. route, which was greater than 2.21 PFU of JEV/ZIKV. Conclusions:The D393E mutation of ZIKV envelope protein in JEV/ZIKA chimeric virus decreases the neurovirulence of the virus in mice.
