重组登革病毒1-4型包膜蛋白EDIII的融合表达和免疫学特性研究

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Abstract Objective: To construct dengue virus envelope protein EDIII tetravalent recombinant vaccines and to analyze its immunogenicity. Methods: Firstly, the gene sequences encoding dengue virus type 1-4 envelope protein EDIII were analyzed by bioinformatics methods, and the representative DENV1-4 EDIII was selected and connected by GS linker to obtain chimeric EDIII; Then, the chimeric gene coding EDIII was inserted into the multiple cloning site of the expression vector pColdIII to constructe the recombinant vector pColdIII-EDIII. The constructed pColdIII-EDIII plasmid was transformed into E.coli BL21 cells and induced by IPTG. The recombinant protein EDIII was purified by Ni-NTA affinity chromatography and confirmed by SDS-PAGE and Western blot analysis. The purified protein was used to immune BABL/c mice. After boosting twice, to detect the antibody titers in serum and the secretion of cytokines by ELISA to evaluate the immune response. Results: The mice immunized with EDIII could induce specific antibodies and the antibody titers was 1:40 000. The mouse spleen lymphocytes stimulated by antigen could produce cytokine including IFN-r, IL-4, IL-10, which were significantly higher in the experimental group. Conclusion: Our studies indicates that dengue virus envelope protein EDIII tetravalent vaccine can induce specific humoral and cellular immune response, and give priority to with the humoral immune response, which laid the foundation for the research of a new type of chimeric tetravalent vaccine in the future.

Key words： dengue virus envelope protein fusion expression tetravalent vaccine

Received: 23 December 2015

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	REN Shoufeng
	LIU Wenquan
	CAO Guomei
	LIANG Shaohui
	PAN Changwang

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2016/V46/I4/235

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