Virologica Sinica

, Volume 22, Issue 1, pp 34–40 | Cite as

Rescue and preliminary application of a recombinant newcastle disease virus expressing green fluorescent protein gene

  • Shun-lin Hu
  • Qin Sun
  • Qu-zhi Wang
  • Yu-liang Liu
  • Yan-tao Wu
  • Xiu-fan Liu
Article
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Accepted:

Abstract

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain, seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI, which contained the full-length cDNA of the NDV ZJI strain. The pNDV/ZJI, with three helper plasmids, pCIneoNP, pCIneoP and pCIneoL, were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase. After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock, an infectious NDV ZJI strain was successfully rescued. Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP. After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells, the recombinant NDV, NDV/ZJIGFP, was rescued. Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection, indicating that the GFP gene was expressed at a relatively high level. NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route. Four days post-infection, strong green fluorescence could be detected in the kidneys and tracheae, indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

Key words

Newcastle disease virus (NDV) Green fluorescent protein (GFP) Rescue Expression 

CLC number

S831.7 
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Copyright information

© Wuhan Institute of Virology, CAS and Springer Berlin Heidelberg 2007

Authors and Affiliations

  • Shun-lin Hu
    • 1
  • Qin Sun
    • 1
  • Qu-zhi Wang
    • 1
  • Yu-liang Liu
    • 1
  • Yan-tao Wu
    • 1
  • Xiu-fan Liu
    • 1
  1. 1.Key Laboratory for Animal Infectious Diseases of Ministry of AgricultureYangzhou UniversityYangzhouChina

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