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Serial Expression of the Truncated Fragments of the Nucleocapsid Protein and Glycoprotein of Crimean-Congo Hemorrhagic Fever Virus and Identification of the Epitope Region

Author: WeiPengFei
Tutor: ZhangFuChun；ZhangYuJiang
School: Xinjiang University
Course: Zoology
Keywords: Crimean - Congo hemorrhagic fever virus Nucleoprotein Glycoprotein Epitope Appraisal
CLC: R373
Type: Master's thesis
Year: 2009
Downloads: 72
Quote: 0
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Abstract


Crimean - Congo haemorrhagic fever (Crimean-Congo hemorrhagic fever, CCHF) is a presence in China's Xinjiang Tarim Basin and Junggar Basin, northern Russia, the Middle East, southern Eurasia and Africa, sub-Saharan potent natural foci of disease , the incidence has obvious seasonal and local, the average mortality rate in the 10% -50%. The pathogen of the disease is a tick-borne Crimean - Congo hemorrhagic fever virus (CCHFV). The virus belongs to the the Bunia virus Conero virus, is a single-stranded negative-strand RNA virus. Currently, there is no effective means for the prevention and treatment of CCHF. In this study, recombinant DNA technology on YL04057 strains of S segment of the Crimean - Congo hemorrhagic fever virus nucleocapsid protein gene of (nucleocapsid Protein, NP) coding region sequence and a series of sub-fragment was cloned into the prokaryotic expression vector, E. coli expression were purified using anti-Crimean - Congo hemorrhagic fever virus polyclonal antibody and two monoclonal antibodies (14B7 and 43E5) epitopes of the expression product testing and analysis. The results show that containing amino acids 235-305 of the NP protein expression of three protein fragments can be anti-Crimean - Congo hemorrhagic fever virus polyclonal antibody detection, can also be reacted with 2 monoclonal antibodies. Showed that of two mAb 14B7 and 43E5 Antibody against epitope positioned in the 235-305 amino acid region of the NP protein, this region is also the NP protein is a highly conserved amino acid region. The same time, we sub-glycoprotein (glycoprotein GP) M fragment coding region fragment was cloned into the prokaryotic expression vector and expressed in E. coli were purified with polyclonal anti-Crimean - Congo hemorrhagic fever virus anti-rabbit antibodies against each of the expression products for the detection and analysis of the epitope. The results showed that the recombinant sub-glycoprotein with rabbit antiserum not. The CCHFV glycoprotein complex structure, and prokaryotic expression systems do not have the complete translation of the modification function. Since the laboratory prepared antiserum for nucleoprotein, the lack of specific detection of glycoproteins. Therefore, it is necessary to establishment of the the CCHFV glycoprotein eukaryotic expression system, glycoprotein and anti-serum, in order to carry out the glycoprotein. Study to determine the Crimean - Congo hemorrhagic fever virus structural protein antigenic characteristics will help Crimean - Congo hemorrhagic fever virus infection and immune mechanisms elucidated, will also provide a theoretical basis for its subunit vaccine.

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CLC: > Medicine, health > Basic Medical > Medical Microbiology ( pathogenic bacteriology,pathogenic microbiology ) > Human Virology ( pathogenic virus)
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