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| Clone, Expression and Immunological Analysis of Type Ⅱ Dengue Virus NS1 Protein |
| LI Duo1, YANG LI-juan2, ZHAO Yu-jiao1, PAN Yue1, CHEN Jun-ying1, FU Juan-juan1, HUANG Xing-wei1, QUE Li-juan1, SUN Qiang-ming1 |
1. Yunan Key laboratory of Vaccine Research Development on Severe Infectious Diseases, Institute of Medical Biology, Chinese Academy of Medical Sciences & Peking Union Medical Colege, Kunming 65018, China;
2. The 2nd Affiliated Hospital of Kunming Medical University, Kunming 650101, China |
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Abstract Objective: To express and prepare the recombinant type II dengue virus nonstructural protein NS1, and study its immunogenicity. Methods: Type Ⅱ Dengue virus NS1 gene were cloned into plasmid pUC57 after codon-optimizing and gene synthesizing. Then the NS1 gene were subcloned into expression vector pET28a. The NS1 protein were inducing expressed by adding IPTG followed by immunoblot identification. Recombinant NS1 protein were renaturated through dialysis and then purified. NS1 polyclonal antibody were preparation by animal immunization. The titer and immunogenicity were detected by indirect enzyme-linked immunological method. Results: The recombinant type II dengue virus nonstructural protein NS1 were successfully expressed, renaturated and purified. It's proved that the recombinant NS1 protein showed good immunogenicity by Western blotting and mice immunization. Conclusion:The recombinant NS1 protein has good immunogenicity, and it's a good beginning for NS1 monoclonal antibody preparation and the development of dengue virus detection kit.
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Received: 20 June 2014
Published: 25 September 2014
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Cite this article:
LI Duo, YANG LI-juan, ZHAO Yu-jiao, PAN Yue, CHEN Jun-ying, FU Juan-juan, HUANG Xing-wei, QUE Li-juan, SUN Qiang-ming. Clone, Expression and Immunological Analysis of Type Ⅱ Dengue Virus NS1 Protein. China Biotechnology, 2014, 34(9): 4-8.
URL:
https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20140902 OR https://manu60.magtech.com.cn/biotech/Y2014/V34/I9/4
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