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

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• 作者：Jianzhong Wang (1)
  Chunfeng Wang (2)
  Na Feng (3)
  Hualei Wang (3)
  Xuexing Zheng (3)
  Songtao Yang (3)
  Yuwei Gao (3)
  Xianzhu Xia (3)
  Renfu Yin (1)
  Xiufan Liu (4)
  Shunlin Hu (4)
  Chan Ding (5)
  Shengqing Yu (5)
  Yanlong Cong (1)
  Zhuang Ding (1)
  
  1. Laboratory of Infectious Diseases ; College of Veterinary Medicine ; Jilin University ; Changchun ; 130062 ; China
  2. Engineering Research Center of Jilin Province for Animals Probiotics ; College of Animal Science and Technology ; Jilin Agricultural University ; Changchun ; 130118 ; China
  3. Institute of Military Veterinary Medicine ; Academy of Military Medical Science ; Changchun ; 130122 ; China
  4. Animal Infectious Disease Laboratory ; College of Veterinary Medicine ; Yangzhou University ; Yangzhou ; 225009 ; Jiangsu ; China
  5. Shanghai Veterinary Research Institute ; Chinese Academy of Agricultural Sciences ; Shanghai ; 200241 ; China
  
• 关键词：NDV ; Genotype VII ; Reverse genetics ; RNA polymerase II
• 刊名：Virus Genes
• 出版年：2015
• 出版时间：February 2015
• 年：2015
• 卷：50
• 期：1
• 页码：152-155
• 全文大小：1,585 KB
• 参考文献：1. Senna, D, Alexander, DJ (2008) Newcastle disease, other avian paramyxoviruses, and pneumovirus infections. Diseases of Poultry. Iowa State University Press, Ames
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• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Biomedicine
  Medical Microbiology
  Virology
  Plant Sciences
  
• 出版者：Springer Netherlands
• ISSN：1572-994X

文摘

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.