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Over-expression of fHbp in Arabdopsis for development of meningococcal serogroup B subunit vaccine

文献类型： 外文期刊

作者： Xu, Nuo 1 ; Wang, Yunpeng 2 ; Ma, Jisheng 4 ; Jin, Libo 3 ; Xing, Shaochen 2 ; Jiang, Chao 3 ; Li, Xiaokun 1 ;

作者机构： 1.Jilin Univ, Coll Life Sci, Changchun 130021, Peoples R China

2.Jilin Acad Agr Sci, Agro Biotechnol Res Inst, Changchun, Peoples R China

3.Wenzhou Univ, Wenzhou, Peoples R China

4.Wenzhou Med Univ, Sch Pharmaceut Sci, Zhejiang Key Lab Biotechnol & Pharmaceut Engn, Wenzhou, Peoples R China

5.Wenzhou Med Univ, Sch Pharmaceut Sci, Zhejiang Key Lab Biotechnol & Pharmaceut Engn, Wenzhou, Peoples R Chin

关键词： Arabidopsis thaliana;fHbp;Neisseria meningitides serogroup B (MenB);Plant-derived subunit vaccine

期刊名称：BIOTECHNOLOGY JOURNAL （ 影响因子：4.677； 五年影响因子：4.596 ）

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收录情况： SCI

摘要： Due to lack of commercial vaccine against the serogroup B (MenB) of Neisseria meningitides, the incidence of meningococcal disease remains high. To solve the issue, transgenic plants are used as bioreactors to produce a plant-derived fHbp subunit vaccine. In this study, the fHbp gene was optimized according to the codon usage bias of Arabidopsis thaliana, synthesized artificially, cloned into an expression vector, driven by a seed-specific promoter, and introduced into A. thaliana by Agrobacterium-mediated floral-dip transformation. Transgenic plants were identified by glufosinate selection, quickstix strips for PAT/bar tests and PCR analysis. The five plants showing higher expression of recombinant fHbp were screened through indirect ELISA. Southern blot analysis showed that the transgenic line rHF-22 had a single-copy integration and the highest expression of fHbp. Recombinant fHbp was purified from seeds of rHF-22 by nitrilotriacetic acid-mediated affinity chromatography, and the purity was 82.5%. BALB/c mice were tested for fHbp vaccine protection from lethal MenB infection, and the relative percent survival was found to be 80%. This study indicates that the recombinant fHbp produced from seeds of rHF-22 is a potential candidate for commercial MenB vaccine. It also provides a reference for safe, cheap and large-scale production of other plant-made vaccines.

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