Serial expression of the truncated fragments of the nucleocapsid protein of CCHFV and identification of the epitope region.
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.
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China.
Peng-fei
PF
Wei
Yan-jun
YJ
Luo
Tian-xian
TX
Li
Hua-lin
HL
Wang
Zhi-hong
ZH
Hu
Fu-chun
FC
Zhang
Yu-jiang
YJ
Zhang
Fei
F
Deng
Su-rong
SR
Sun
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000914:Antibodies, Viral / Q000276:immunology; D015153:Blotting, Western; D018604:Epitope Mapping / Q000379:methods*; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D003225:Hemorrhagic Fever Virus, Crimean-Congo / Q000235:genetics / Q000276:immunology*; D050505:Mutant Proteins / Q000235:genetics / Q000276:immunology*; D019590:Nucleocapsid Proteins / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D017384:Sequence Deletion
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000939:Epitopes; D050505:Mutant Proteins; D019590:Nucleocapsid Proteins; D011994:Recombinant Proteins