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Volume 25 Issue 1
February 2010
    Citation: Peng-fei WEI, Yan-jun LUO, Tian-xian LI, Hua-lin WANG, Zhi-hong HU, Fu-chun ZHANG, Yu-jiang ZHANG, Fei DENG, Su-rong SUN. Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region .VIROLOGICA SINICA, 2010, 25(1) : 45-51.  http://dx.doi.org/10.1007/s12250-010-3067-7
    shu

    Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region

    cstr: 32224.14.s12250-010-3067-7
    • 1.

      Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China

    • 2.

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

    • 3.

      The Center for Disease Control and Prevention of Xinjiang Uygur Autonomous Region, Urumqi 830002, China

    • Corresponding author: Fei DENG, df@wh.iov.cn
      Su-rong SUN, sunsr@xju.edu.cn
    • Received Date: 30 April 2009
      Accepted Date: 24 June 2009
      Available online: 01 February 2010

      Fund Project: National Key Technologies R & D Program of China during the 10th Five-Year Plan Period 2003BA712A08-03The NSFC Grant 30860255The Knowledge Innovation Program of the Chinese Academy of Sciences KSCX2-YW-N-065The Foundation scientific and technological project from MOST 2007FY210700

    • The Crimean-congo hemorrhagic fever virus (CCHFV) is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections. In this study, expression vectors carrying series truncated fragments of the NP ( nucleocapsid protein ) gene from the S fragment of CCHFV strain YL04057 were constructed. The recombinant proteins were expressed in E.coli and purified for detection. The antigenic of the truncated fragments of NP was detected with a polyclonal serum (rabbit) and 2 monoclonal (mAbs)（14B7 and 43E5）against CCHFV by Western-blot analyses. The results showed that the three expressed constructs, which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum. The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein
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      1. Calisher C H. 1996. History, classification and taxonomy of viruses in the family Bunyaviridae. In: The Bunyaviridae (Elliott R M. ed.), New York: Plenum Press, p1-17.

      2. Deyde V M, Khristova M L, Rollin P E, et al. 2006. Crimean-Congo Hemorrhagic Fever Virus Genomics and Global Diversity. J Vriol, 80 (17): 8834-8842.

      3. Fack F, Hugle-Dorr B, Song D, et al. 1997. Epitope mapping by phage display: Random versus gene-fragment libraries. J Immunol Metho, 206 (1-2), 43-52.
          doi: 10.1016/S0022-1759(97)00083-5

      4. Feng C H. 2004. Experimental Observation on Recog-nition of Viral Protein by Monoclonal Antibodies to XHF Virus. Endemic Diseases Bulletin, 19, 94-96.

      5. Han L, Tang Q, Zhao X Q, et al. 2001. Serologic studies of Xinjiang hemorrhagic fever in Bachu county. Chin J Epidemiol, 23, 179-181.

      6. Hoogstraal H. 1979. The epidemiology of tick borne Crimean-Congo hemorrhagic fever in Asia, Europe, and Africa. J Med Entomol, 15, 307-417.
          doi: 10.1093/jmedent/15.4.307

      7. Larsen J E, Lund O, Nielsen M. 2006. Improved method for predicting linear B-cell epitopes. Immunome Res, 24, 2: 2.

      8. Saijo M, Tang Q, Niikura M, et al. 2002. Recombinant Nucleoprotein-Based Enzyme-Linked Immunosorbent Assay for Detection of Immunoglobulin G Antibodies Crimean-Congo Hemorrhagic Fever Virus. J Clin MicroBiol, 40 (5): 1587-1591.
          doi: 10.1128/JCM.40.5.1587-1591.2002

      9. Saijo M, Tang Q, Shimayi B, et al. 2005. Antigen-capture enzyme-linked immunosorbent assay for the diagnosis of crimean-congo hemorrhagic fever using a novel mono-clonal antibody. J Med Virol, 77: 83-88.
          doi: 10.1002/(ISSN)1096-9071

      10. Schmaljohn C S, Hooper J W. 2001. Bunyaviridae: the viruses and their replication. In: Fields virology, 4th ed. Philadelphia: Lippincott-Raven Publishers, p1581-1602.

      11. Shepherd A J, Swanepoel R, Shepherd S P, et al. 1985. A nosocomial outbreak of Crimean-Congo haemorrhagic fever at Tygerberg hospital. Part 5. Virological and serological observations. S Afr Med J, 68 (10), 733-736.

      12. Swanepoel R, Shepherd A J, Leman P A, et al. 1987. Epidemiological and clinical features of Crimean-Congo haemorrhagic fever in southern Africa. Am J Trop Med Hyg, 36 (1), 120-132.
          doi: 10.4269/ajtmh.1987.36.120

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      Serial Expression of the Truncated Fragments of the Nucleocapsid Protein of CCHFV and Identification of the Epitope Region

        Corresponding author: Fei DENG, df@wh.iov.cn
        Corresponding author: Su-rong SUN, sunsr@xju.edu.cn
      • 1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China
      • 2. State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China
      • 3. The Center for Disease Control and Prevention of Xinjiang Uygur Autonomous Region, Urumqi 830002, China
      • Received Date: 30 April 2009
      • Accepted Date: 24 June 2009
      Fund Project:  National Key Technologies R & D Program of China during the 10th Five-Year Plan Period 2003BA712A08-03The NSFC Grant 30860255The Knowledge Innovation Program of the Chinese Academy of Sciences KSCX2-YW-N-065The Foundation scientific and technological project from MOST 2007FY210700

      Abstract: The Crimean-congo hemorrhagic fever virus (CCHFV) is a geographically widespread fatal pathogen. Identification of the epitope regions of the virus is important for the diagnosis and epidemiological studies of CCHFV infections. In this study, expression vectors carrying series truncated fragments of the NP ( nucleocapsid protein ) gene from the S fragment of CCHFV strain YL04057 were constructed. The recombinant proteins were expressed in E.coli and purified for detection. The antigenic of the truncated fragments of NP was detected with a polyclonal serum (rabbit) and 2 monoclonal (mAbs)（14B7 and 43E5）against CCHFV by Western-blot analyses. The results showed that the three expressed constructs, which all contained the region 235AA to 305AA could be detected by mAbs polyclonal serum. The results suggest that region 235-305 aa of NP is a highly antigenic region and is highly conserved in the NP protein

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