Genetically stable reporter virus, subgenomic replicon and packaging system of duck Tembusu virus based on a reverse genetics system

Yu He; Peng Liu; Tao Wang; Yuanyuan Wu; Xiao Lin; Mingshu Wang; Renyong Jia; Dekang Zhu; Mafeng Liu; Xinxin Zhao; Qiao Yang; Ying Wu; Shaqiu Zhang; Yunya Liu; Ling Zhang; Yanling Yu; Leichang Pan; Shun Chen; Anchun Cheng

doi:10.1016/j.virol.2019.05.003

Genetically stable reporter virus, subgenomic replicon and packaging system of duck Tembusu virus based on a reverse genetics system

Virology. 2019 Jul:533:86-92. doi: 10.1016/j.virol.2019.05.003. Epub 2019 May 18.

Authors

Yu He¹, Peng Liu¹, Tao Wang¹, Yuanyuan Wu¹, Xiao Lin¹, Mingshu Wang², Renyong Jia², Dekang Zhu³, Mafeng Liu², Xinxin Zhao², Qiao Yang², Ying Wu², Shaqiu Zhang², Yunya Liu¹, Ling Zhang¹, Yanling Yu¹, Leichang Pan¹, Shun Chen⁴, Anchun Cheng⁵

Affiliations

¹ Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
² Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
³ Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China.
⁴ Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. Electronic address: [email protected].
⁵ Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Research Center of Avian Disease, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China; Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan, 611130, China. Electronic address: [email protected].

PMID: 31136895
DOI: 10.1016/j.virol.2019.05.003

Abstract

Duck Tembusu virus (DTMUV) is a novel flavivirus that has caused an outbreak of severe duck egg-drop syndrome since 2010. It has spread rapidly to other avian species, causing enormous economic loss. In the present study, we generated a reporter virus expressing NanoLuc luciferase, which was stable after 10 rounds of continuous propagation without reporter gene deletion. Moreover, we generated two types of replicons driven by the T7 promoter or CMV promoter, both of which worked well in BHK21 cells. Furthermore, we developed the first packaging system for DTMUV by co-transfection into BHK21 cells of a replicon (containing mature C) and a plasmid encoding C₁₆-prM-E, which resulted in the production of single round infectious particles (SRIPs). We also generated a packaging cell line for DTMUV to produce SRIPs. We believe that these multicomponent platform tools are important for DTMUV pathogenesis research and novel vaccine development.

Keywords: Duck Tembusu virus; Packaging cell line; Packaging system; Reporter virus; Subgenomic replicon.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Ducks
Flavivirus / genetics*
Flavivirus / physiology
Flavivirus Infections / veterinary*
Flavivirus Infections / virology
Genes, Reporter
Luciferases / genetics
Luciferases / metabolism
Poultry Diseases / virology*
Replicon*
Reverse Genetics

Substances

Luciferases

Supplementary concepts

Tembusu virus