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Development of a reverse genetics system based on RNA polymerase II for Newcastle disease virus genotype VII

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Abstract

Newcastle disease virus (NDV) has only a single serotype but diversified genotypes. Genotype VII strains are the prevalent currently circulating genotype worldwide, and in particular, these strains cause outbreaks in waterfowl. In this study, a reverse genetics system for highly virulent NDV isolated from goose flocks was developed independent of conventional T7 RNA polymerase. Infectious virus was successfully generated by an RNA polymerase II promoter to drive transcription of the full-length virus antigenome. A green fluorescent protein (GFP)-expressing virus was generated by inserting an additional transcription cassette coding for the enhanced GFP between the P and M genes of the genome. The expression of GFP was confirmed by western blotting and fluorescence microscopy. The replication kinetics and pathogenicity of the recombinant viruses are indistinguishable from the parental wild-type virus. This reverse genetics system will provide a powerful tool for the analysis of goose-origin NDV dissemination and pathogenesis, as well as preparation for genotype-matched NDV attenuated vaccines.

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Acknowledgments

The research was supported by the National Natural Scientific Fund (31272561), Chinese Special Fund for Agro-scientific Research in the Public Interest (201303033), and one funding from Jilin Provincial Science & Technology department (20140520171JH).

Conflict of interest

The authors have declared that no competing interests exist

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Authors and Affiliations

  1. Laboratory of Infectious Diseases, College of Veterinary Medicine, Jilin University, Changchun, 130062, China

    Jianzhong Wang, Renfu Yin, Yanlong Cong & Zhuang Ding

  2. Engineering Research Center of Jilin Province for Animals Probiotics, College of Animal Science and Technology, Jilin Agricultural University, Changchun, 130118, China

    Chunfeng Wang

  3. Institute of Military Veterinary Medicine, Academy of Military Medical Science, Changchun, 130122, China

    Na Feng, Hualei Wang, Xuexing Zheng, Songtao Yang, Yuwei Gao & Xianzhu Xia

  4. Animal Infectious Disease Laboratory, College of Veterinary Medicine, Yangzhou University, Yangzhou, 225009, Jiangsu, China

    Xiufan Liu & Shunlin Hu

  5. Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, China

    Chan Ding & Shengqing Yu

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  1. Jianzhong Wang
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  2. Chunfeng Wang
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  3. Na Feng
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  4. Hualei Wang
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  7. Yuwei Gao
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  8. Xianzhu Xia
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  9. Renfu Yin
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  10. Xiufan Liu
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  11. Shunlin Hu
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  12. Chan Ding
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  13. Shengqing Yu
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  14. Yanlong Cong
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  15. Zhuang Ding
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Corresponding authors

Correspondence to Yanlong Cong or Zhuang Ding.

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Wang, J., Wang, C., Feng, N. et al. Development of a reverse genetics system based on RNA polymerase II for Newcastle disease virus genotype VII. Virus Genes 50, 152–155 (2015). https://doi.org/10.1007/s11262-014-1137-x

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  • DOI: https://doi.org/10.1007/s11262-014-1137-x

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