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Construction of a minigenome rescue system for Newcastle disease virus strain Italien

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Abstract

Newcastle disease virus (NDV) Italien, a velogenic strain, is an oncolytic virus that is considered to be a potential agent for antitumor viral therapy. We constructed three helper plasmids expressing the NP, P and L genes of NDV Italien based on the eukaryotic expression plasmid pcDNA3.1(+). The minigenome consisting of the 3′ leader and 5′ trailer regions of NDV Italien flanking a reporter gene encoding firefly luciferase was constructed to examine the efficacy of the three helper plasmids in viral genome replication and transcription. After co-transfection of BSR-T7/5 cells with the three helper plasmids and the minigenome plasmid, replication of minigenome RNA was evaluated by determining luciferase activity. In the minigenome rescue system, expression of the reporter gene was detected. Our results indicate that the three proteins NP, P, and L are correctly expressed and can assemble into a functional ribonucleoprotein complex that effectively directs the transcription of minigenome RNA.

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Acknowledgments

This study was supported by the National Natural Science Foundation of China (no. 30671196 and 30870944). We thank Dr. Ben Peeters for his discussion during the construction of plasmids and revision of this manuscript. We thank Prof. Karl-Klaus Conzelmann for providing us the BSR-T7/5 cells, and Prof. Volker Schirrmacher for the NDV strain Italien and rabbit anti-NDV serum.

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Authors and Affiliations

  1. Cell Engineering Research Centre and Department of Cell Biology, State Key Laboratory of Cancer Biology, Fourth Military Medical University, No.17 Changle West Road, Xi’an, 710032, China

    Hao Feng, Ding Wei, Gang Nan, Zhi-Nan Chen & Huijie Bian

  2. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, 150001, China

    Shang-Jin Cui

Authors
  1. Hao Feng
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  2. Ding Wei
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  3. Gang Nan
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  4. Shang-Jin Cui
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  5. Zhi-Nan Chen
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  6. Huijie Bian
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Corresponding authors

Correspondence to Zhi-Nan Chen or Huijie Bian.

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H. Feng and D. Wei contributed equally to this work.

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Feng, H., Wei, D., Nan, G. et al. Construction of a minigenome rescue system for Newcastle disease virus strain Italien. Arch Virol 156, 611–616 (2011). https://doi.org/10.1007/s00705-010-0898-3

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  • DOI: https://doi.org/10.1007/s00705-010-0898-3

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