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Volume 24 Issue 6
December 2009
    Citation: Ping-gang MING, Ying HUANG, Qing TANG, Jia-liang DU, Xiao-yan TAO, Jia-xin YAN, Rong-liang HU. Construction and Identification of the Helper Plasmids for Reverse Genetic System of Rabies Virus Street Strain .VIROLOGICA SINICA, 2009, 24(6) : 559-565.  http://dx.doi.org/10.1007/s12250-009-3039-y
    shu

    Construction and Identification of the Helper Plasmids for Reverse Genetic System of Rabies Virus Street Strain

    cstr: 32224.14.s12250-009-3039-y
    • 1.

      State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (IVDC, China CDC), Beijing 100052, China

    • 2.

      Department of Genetic Engineering, Wuhan Institute of Biological Products, Wuhan 430060, China

    • 3.

      Laboratory of Epidemiology, Veterinary Institute, Academy of Military Medical Sciences, Changchun 130062, China

    • Corresponding author: Qing TANG, qtang04@sina.com
    • Received Date: 17 March 2009
      Accepted Date: 08 June 2009
      Available online: 01 December 2009

      Fund Project: Major Program of the National Natural Science Foundation of China 30630049National High Technology Research and Development Program of China 2006AA02Z110National High Technology Research and Development Program of China 2007AA02Z402

    • To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.
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      1. Ai J, Lin B Z, Guo X F. 2006. Rescue of HEP-Flury strain of rabies virus from cloned cDNA. Prog Veter Med, 27 (2): 99-100. (in Chinese)

      2. Conzelmann K K, Schnell M. 1994. Rescue of synthetic genomic RNA analogs of Rabies virus by plasmid-encoded protein. J Virol, 68 (2): 713-719.

      3. Guo X F, Fu Z F. 2006. Moving the glycoprotein gene of rabies virus to promoter-proximal position and the generation of the virus. J South China Agr Univ, 27 (1): 104-106. (in Chinese).

      4. Huang Z, Krishnamurthy S, Panda A, et al.Newcastle Disease Virus V Protein Is Associated with Viral Pathogenesis and Functions as an Alpha Interferon Antagonist. J Virol, 77 (16): 8676-8685.

      5. Inoue K, Shoji Y, Kurane I, et al. 2003. An improved method for recovering rabies virus from cloned cDNA. J Virol Methods, 107 (2): 229-236.
          doi: 10.1016/S0166-0934(02)00249-5

      6. Ito N, Takayama M, Yamada K, et al. 2001. Rescue of rabies virus from cloned cDNA and identification of the pathogenicity-related gene: glycoprotein gene is associated with virulence for adult mice. J Virol, 75 (19): 9121-9128.
          doi: 10.1128/JVI.75.19.9121-9128.2001

      7. Ming P G, Huang Y, Tang Q, et al. 2009. Construction and analysis of full-length cDNA of rabies virus street strain. Chin J Virol, 25 (1): 17-22. (in Chinese)

      8. Ming P G, Du J L, Tang Q, et al. 2009. Molecular characterization of the complete genome of a street rabies virus isolated in China. Virus Res, 143 (1): 6-14.
          doi: 10.1016/j.virusres.2009.02.014

      9. Morimoto K, Foley H D, McGettigan J P, et al. 2000. Reinvestigation of the role of the rabies virus glycoprotein in viral pathogenesis using a reverse genetics approach. J Neurovirol, 6 (5): 373-381.
          doi: 10.3109/13550280009018301

      10. Schnell M J, Mebatsion T, Conzelmann K K. 1994. Infectious rabies viruses from cloned cDNA. EMBO J, 13 (18): 4195-4203.

      11. Tordo N, Poch O, Ermine A, et al. 1986. Walking along the rabies genome: is the large G-L intergenic region a remnant gene? Proc Natl Acad Sci USA, 83 (11): 3914-3918.
          doi: 10.1073/pnas.83.11.3914

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      Construction and Identification of the Helper Plasmids for Reverse Genetic System of Rabies Virus Street Strain

        Corresponding author: Qing TANG, qtang04@sina.com
      • 1. State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention (IVDC, China CDC), Beijing 100052, China
      • 2. Department of Genetic Engineering, Wuhan Institute of Biological Products, Wuhan 430060, China
      • 3. Laboratory of Epidemiology, Veterinary Institute, Academy of Military Medical Sciences, Changchun 130062, China
      • Received Date: 17 March 2009
      • Accepted Date: 08 June 2009
      Fund Project:  Major Program of the National Natural Science Foundation of China 30630049National High Technology Research and Development Program of China 2006AA02Z110National High Technology Research and Development Program of China 2007AA02Z402

      Abstract: To obtain the helper plasmids for a reverse genetics system of rabies virus, the cDNAs of the complete open reading frames of the N, P, G, and L genes of rabies street virus stain HN10 were each cloned into expression vector pVAX1, These four plasmids were identified by restriction enzyme digestion and gene sequencing. The plasmid encoding the N protein was selected to determine the expression effect of these plasmids in NA cells. The results showed that the helper plasmids for a reverse genetics system of rabies street virus strain HN10 had been successfully constructed.

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