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Development and optimization of a DNA-based reverse genetics systems for epizootic hemorrhagic disease virus

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Abstract

Epizootic hemorrhagic disease virus (EHDV) is a member of the genus Orbivirus, family Reoviridae, and has a genome consisting of 10 linear double-stranded (ds) RNA segments. The current reverse genetics system (RGS) for engineering the EHDV genome relies on the use of in vitro-synthesized capped viral RNA transcripts. To obtain more-efficient and simpler RGSs for EHDV, we developed an entirely DNA (plasmid or PCR amplicon)-based RGS for viral rescue. This RGS enabled the rescue of infectious EHDV from BSR-T7 cells following co-transfection with seven helper viral protein expression plasmids and 10 cDNA rescue plasmids or PCR amplicons representing the EHDV genome. Furthermore, we optimized the DNA-based systems and confirmed that some of the helper expression plasmids were not essential for the recovery of infectious EHDV. Thus, DNA-based RGSs may offer a more efficient method of recombinant virus recovery and accelerate the study of the biological characteristics of EHDV and the development of novel vaccines.

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Funding

This study was supported by the National Key R&D Program of China (2017YFD0500903) and the Central Public-Interest Scientific Institution Basal Fund (1610302016008).

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Authors and Affiliations

  1. The Key Laboratory of Veterinary Public Health, Ministry of Agriculture, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, 150069, China

    Yunze Guo, Qingyuan Xu, Donglai Wu, Zhigao Bu & Encheng Sun

  2. Department of Veterinary Pathology, Laboratory of Clinical Diagnosis and Treatment Technology in Animal Disease, Ministry of Agriculture, College of Veterinary Medicine, Inner Mongolia Agricultural University, Zhaowuda Road 306, 010018, Hohhot, Inner Mongolia, China

    Yunze Guo

  3. Department of Biochemistry, Genetics and Microbiology, University of Pretoria, Pretoria, 0002, South Africa

    Jakobus M. Pretorius & Jacques Theron

Authors
  1. Yunze Guo
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  2. Jakobus M. Pretorius
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  3. Qingyuan Xu
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  4. Donglai Wu
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  5. Zhigao Bu
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  6. Jacques Theron
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  7. Encheng Sun
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Correspondence to Encheng Sun.

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The authors declare that there are no conflicts of interest.

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Guo, Y., Pretorius, J.M., Xu, Q. et al. Development and optimization of a DNA-based reverse genetics systems for epizootic hemorrhagic disease virus. Arch Virol 165, 1079–1087 (2020). https://doi.org/10.1007/s00705-020-04583-w

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  • DOI: https://doi.org/10.1007/s00705-020-04583-w

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