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重组表达 mCherry 报告蛋白的西尼罗河病毒的构建。

Development of recombinant West Nile virus expressing mCherry reporter protein.

机构信息

Laboratory of Public Health, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

Laboratory of Public Health, Faculty of Veterinary Medicine, Hokkaido University, Sapporo, Japan.

出版信息

J Virol Methods. 2023 Jul;317:114744. doi: 10.1016/j.jviromet.2023.114744. Epub 2023 Apr 28.

Abstract

West Nile virus (WNV) is transmitted to humans and animals by a mosquito and enters the central nervous system, leading to lethal encephalitis. Reporter viruses expressing fluorescent proteins enable detection of infected cells in vitro and in vivo, facilitating evaluation of the dynamics of viral infection, and the development of diagnostic or therapeutic methods. In this study, we developed a method for production of a recombinant replication-competent WNV expressing mCherry fluorescent protein. The expression of mCherry was observed in viral antigen-positive cells in vitro and in vivo, but the growth of the reporter WNV was reduced as compared to the parental WNV. The expression of mCherry was stable during 5 passages in reporter WNV-infected culture cells. Neurological symptoms were observed in mice inoculated intracranially with the reporter WNV. The reporter WNV expressing mCherry will facilitate research into WNV replication in mouse brains.

摘要

西尼罗河病毒(WNV)通过蚊子传播给人类和动物,并进入中枢神经系统,导致致命的脑炎。表达荧光蛋白的报告病毒使我们能够在体外和体内检测感染细胞,从而有助于评估病毒感染的动态,并开发诊断或治疗方法。在这项研究中,我们开发了一种生产表达 mCherry 荧光蛋白的重组复制型 WNV 的方法。在体外和体内,观察到 mCherry 在病毒抗原阳性细胞中的表达,但与亲本 WNV 相比,报告 WNV 的生长受到了抑制。在报告 WNV 感染的培养细胞中进行 5 次传代时,mCherry 的表达保持稳定。用报告 WNV 经颅接种的小鼠出现了神经症状。表达 mCherry 的报告 WNV 将有助于研究 WNV 在小鼠大脑中的复制。

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