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Volume 26 Issue 5
October 2011
    Citation: Zhao-rui Zhou, Man-li Wang, Fei Deng, Tian-xian Li, Zhi-hong Hu, Hua-lin Wang. Production of CCHF Virus-Like Particle by a Baculovirus-Insect Cell Expression System .VIROLOGICA SINICA, 2011, 26(5) : 338-346.  http://dx.doi.org/10.1007/s12250-011-3209-6
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    Production of CCHF Virus-Like Particle by a Baculovirus-Insect Cell Expression System

    cstr: 32224.14.s12250-011-3209-6

    • State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China

    • Corresponding author: Man-li Wang, Wangml@wh.iov.cn
    • Received Date: 29 June 2011
      Accepted Date: 31 August 2011
      Available online: 01 October 2011

      Fund Project: The Knowledge Innovation Program of the Chinese Academy of Sciences, Grant KSCX2-YW-N-065National Basic Research Priorities Program of China, Grant 2007National Key Basic Research Program (973 Program), Grant 2010CB530103the Knowledge Innovation Program of the Chinese Academy of Sciences, Grant KSCX2-EWG-8

    • Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family, which is widespread and causes high fatality. The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment. In this research, the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus. Under an electron microscope, Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells. Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation, which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).
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      1. Barr J N, Wertz G W. 2004. Bunyamwera bunyavirus RNA synthesis requires cooperation of 3'-and 5'-terminal sequences. J Virol, 78: 1129-1138.
          doi: 10.1128/JVI.78.3.1129-1138.2004

      2. Chai J J, Xiao C E, Liu Y H, et al. 2004. Report of Investigation on Hemorrhagic Fever in Bachu County, Xinjiang in 1966 Ⅲ. Investigation an Epidemiology of Hemorrhagic Fever in Bachu. Endemic Disease Bulletin (in chinese), 19S: 15-22.

      3. Chinikar S, Ghiasi S M, Hewson R, et al. 2010. Crimean-Congo hemorrhagic fever in Iran and neighboring countries. J Clin Virol, 47: 110-114.
          doi: 10.1016/j.jcv.2009.10.014

      4. Deng F, Wang R R, Fang M G, et al. 2007. Proteomics analysis of Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus identified two new occlusion-derived virus-associated proteins, HA44 and HA100. J Virol, 81: 9377-9385.
          doi: 10.1128/JVI.00632-07

      5. Deyde V M, Khristova M L, Rollin P E, et al. 2006. Crimean-Congo hemorrhagic fever virus genomics and global diversity. J Virol, 80: 8834-8842.
          doi: 10.1128/JVI.00752-06

      6. Elevli M, Ozkul A A, Civilibal M, et al. 2010. A newly identified Crimean-Congo hemorrhagic fever virus strain in Turkey. Int J Infect Dis, 14 Suppl (3): e213-216.

      7. Elliott R M. 1990. Molecular biology of the Bunyaviridae. J Gen Virol, 71: 501-522.
          doi: 10.1099/0022-1317-71-3-501

      8. Ergonul O. 2006. Crimean-Congo haemorrhagic fever. Lancet Infect Dis, 6: 203-214.
          doi: 10.1016/S1473-3099(06)70435-2

      9. Ergonul O. 2008. Treatment of Crimean-Congo hemorrhagic fever. Antiviral Res, 78: 125-131.
          doi: 10.1016/j.antiviral.2007.11.002

      10. Flick R. 2007. Chapter 4: Molecular Biology of the Crimean-Congo Hemorrhagic Fever Virus. In: Crimean-Congo Hemorrhagic Fever (O Ergonul, C A Whitehouse ed. ). Springer Netherlands, p 35-44.

      11. Frank S A. 2002. Immunology and Evolution of Infectious Disease. Princeton (NJ): Pricenton University Press, p94-108.

      12. Fu T M, Guan L, Friedman A, et al. 1999. Dose dependence of CTL precursor frequency induced by a DNA vaccine and correlation with protective immunity against influenza virus challenge. J Immunol, 162: 4163-4170.

      13. Ghodake R S, Babar R S, Patil S B, et al. 2011. CCHFV-an emerging infectious disease. Pharma science monitor: An international journal of pharmaceutical sciences, 1067-1087.

      14. Hewson R, Chamberlain J, Mioulet V, et al. 2004. Crimean-Congo haemorrhagic fever virus: sequence analysis of the small RNA segments from a collection of viruses world wide. Virus Res, 102: 185-189.
          doi: 10.1016/j.virusres.2003.12.035

      15. King L A, Possee R D. 1992. The baculovirus expression system: A laboratory guide. New York, NY (USA): Chapman & Hall. p158-159.

      16. Kunkel M, Lorinczi M, Rijnbrand R, et al. 2001. Self-assembly of nucleocapsid-like particles from recombinant hepatitis C virus core protein. J Virol, 75: 2119-2129.
          doi: 10.1128/JVI.75.5.2119-2129.2001

      17. Lazo L, Gil L, Lopez C, et al. 2010. Nucleocapsid-like particles of dengue-2 virus enhance the immune response against a recombinant protein of dengue-4 virus. Arch Virol, 155: 1587-1595.
          doi: 10.1007/s00705-010-0734-9

      18. Liu L, Celma C C, Roy P. 2008. Rift Valley fever virus structural proteins: expression, characterization and assembly of recombinant proteins. Virol J, 5: 82.
          doi: 10.1186/1743-422X-5-82

      19. Liu S J, Leng C H, Lien S P, et al. 2006. Immunological characterizations of the nucleocapsid protein based SARS vaccine candidates. Vaccine, 24: 3100-3108.
          doi: 10.1016/j.vaccine.2006.01.058

      20. Lopez C, Gil L, Lazo L, et al. 2009. In vitro assembly of nucleocapsid-like particles from purified recombinant capsid protein of dengue-2 virus. Arch Virol, 154: 695-698.
          doi: 10.1007/s00705-009-0350-8

      21. Ma B J, Wang H. 2000. The hacker of mount Tianshan— Crimean-Congo hemorrhagic fever virus. Foreign medical Sciences (Section of Virology) (in chinese), 7: 8-10.

      22. Midilli K, Gargılı A, Ergonul O, et al. 2007. Imported Crimean-Congo hemorrhagic fever cases in Istanbul. BMC Infectious Diseases, 7: 54.
          doi: 10.1186/1471-2334-7-54

      23. Ozdarendeli A, Canakoglu N, Berber E, et al. 2010. The complete genome analysis of Crimean-Congo hemorrhagic fever virus isolated in Turkey. Virus Res, 147: 288-293.
          doi: 10.1016/j.virusres.2009.11.009

      24. Reguera J, Weber F, Cusack S. 2010. Bunyaviridae RNA polymerases (L-protein) have an N-terminal, influenza-like endonuclease domain, essential for viral cap-dependent transcription. PLoS Pathogens, 6: 1-14.

      25. Saijo M, Qing T, Niikura M, et al. 2002. Recombinant nucleoprotein-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Crimean-Congo hemorrhagic fever virus. J Clin Microbiol, 40: 1587-1591.
          doi: 10.1128/JCM.40.5.1587-1591.2002

      26. Tang Q. 2006. Crimean-Congo hemorrhagic fever virus recently research progress. Chinese J Exp Clin Virol (in chinese), 20: 86-89.

      27. Wang M L, Tan Y, Yin F F, et al. 2008. The F-like protein Ac23 enhances the infectivity of the budded virus of gp64-null Autographa californica multinucleocapsid nucleopolyhedrovirus pseudotyped with baculovirus envelope fusion protein F. J Virol, 82: 9800-9804.
          doi: 10.1128/JVI.00759-08

      28. Wang M L, Tuladhar E, Shen S, et al. 2010. Specificity of baculovirus P6.9 basic DNA-binding proteins and critical role of the C terminus in virion formation. J Virol, 84: 8821-8828.
          doi: 10.1128/JVI.00072-10

      29. Wang R l, Jin N Y, Zhao B, et al. 2005. Expression of the Gene Encoding the N Protein of SARS Virus in E.coli and Its Usd in the Immunization of Hourse. Virologica Sinica (in chinese), 20: 582-585.

      30. Wasmoen T L, Kadakia N P, Unfer R C, et al. 1995. Protection of cats from infectious peritonitis by vaccination with a recombinant raccoon poxvirus expressing the nucleocapsid gene of feline infectious peritonitis virus. Adv Exp Med Biol, 380: 221-228.
          doi: 10.1007/978-1-4615-1899-0

      31. Wei P F, Luo Y J, Li T X, et al. 2010. Serial expression of the truncated fragments of the nucleocapsid protein of CCHFV and identification of the epitope region. Virologica Sinica, 25: 45-51.
          doi: 10.1007/s12250-010-3067-7

      32. Whitehouse C A. 2004. Crimean-Congo hemorrhagic fever. Antiviral Res, 64: 145-160.
          doi: 10.1016/S0166-3542(04)00163-9

      33. Wilson J A, Hart M K. 2001. Protection from Ebola virus mediated by cytotoxic T lymphocytes specific for the viral nucleoprotein. J Virol, 75: 2660-2664.
          doi: 10.1128/JVI.75.6.2660-2664.2001

      34. Xia H, Li P, Yang J, et al 2011. Epidemiological survey of Crimean-Congo hemorrhagic fever virus in Yunnan, China, 2008. Int J Infect Dis, 15: e459-463.
          doi: 10.1016/j.ijid.2011.03.013

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      Production of CCHF Virus-Like Particle by a Baculovirus-Insect Cell Expression System

        Corresponding author: Man-li Wang, Wangml@wh.iov.cn
      • State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
      • Received Date: 29 June 2011
      • Accepted Date: 31 August 2011
      Fund Project:  The Knowledge Innovation Program of the Chinese Academy of Sciences, Grant KSCX2-YW-N-065National Basic Research Priorities Program of China, Grant 2007National Key Basic Research Program (973 Program), Grant 2010CB530103the Knowledge Innovation Program of the Chinese Academy of Sciences, Grant KSCX2-EWG-8

      Abstract: Crimean-Congo Haemorrhagic Fever Virus (CCHFV) is a tick-born virus of the Nairovirus genus within the Bunyaviridae family, which is widespread and causes high fatality. The nucleocapsid of CCHFV is comprised of N proteins that are encoded by the S segment. In this research, the N protein of CCHFV was expressed in insect cells using a recombinant baculovirus. Under an electron microscope, Virus-Like Particles (VLPs) with various size and morphology were observed in cytoplasmic vesicles in the infected cells. Sucrose-gradient purification of the cell lysate indicated that the VLPs were mainly located in the upper fraction after ultracentrifugation, which was confirmed by Western blot analysis and immuno-electron microscopy (IEM).

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