Laboratory Animal and Comparative Medicine ›› 2015, Vol. 35 ›› Issue (3): 195-201.DOI: 10.3969/j.issn.1674-5817.2015.03.004

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Construction of Recombinant Duck Enteritis Virus with HN Gene of Newcastle Disease Virus

LIANG Shu-lin1, LI Hui-xin2, CHEN Hong-yan3, LIU Sheng-wang1,2   

  1. 1. College of Life Science, Northeast Agricultural University, Harbin 150030, China;
    2. Innovation Team of Disease of Avian Respiratory Tract, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China;
    3. Experimental Animal Center, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
  • Received:2014-12-26 Online:2015-06-25 Published:2015-06-25

Abstract: Objective To construct the recombinant duck enteritis virus (DEV) expressing the HN gene of Newcastle disease virus (NDV). Methods The HN gene of NDV was inserted into TK gene of DEV Clone-03 by homologous recombination technology. rDEV TK-EGFP which was constructed as parent virus, by means of cotransfection of the viral genome and transfer vector and plaque purification, the recombinant duck enteritis virus was obtained expression the HN gene of Newcastle disease virus. Results The HN gene of NDV was inserted into DEV genome correctly and expressed effectively on gene and protein level. Replication kinetics and the genetic stability detection of the recombinant virus rDEV TK-HN proved that the titer of rDEV TK-HN decreased slightly, but the trend was consistent with the parent virus replication. After 20 passages in chicken embryo fibroblast and SPF chicken embryos, it showed that the HN gene was of good genetic and expression stability with the passage of the virus. Conclusion By homologous recombination technology and modifying duck enteritis virus genome, the recombinant virus which could carry the HN gene of NDV was constructed and the virus was of good genetic and expression stability.

Key words: Duck enteritis virus (DEV), Newcastle disease virus (NDV), HN gene, Recombinant virus, Genetic modification

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