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Over‐expression of fHbp in Arabdopsis for development of meningococcal serogroup B subunit vaccine
Authors:Nuo Xu  Yunpeng Wang  Jisheng Ma  Libo Jin  Shaochen Xing  Chao Jiang  Xiaokun Li
Institution:1. College of Life Sciences, Jilin University, Changchun, China;2. Wenzhou University, Wenzhou, China;3. Agro-Biotechnology Research Institute, Jilin Academy of Agriculture Science, Changchun, China;4. School of Pharmaceutical Sciences, Zhejiang Key Laboratory of Biotechnology and Pharmaceutical Engineering, Wenzhou Medical University, Wenzhou, China
Abstract: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.
Keywords:Arabidopsis thaliana  fHbp  Neisseria meningitides serogroup B (MenB)  Plant‐derived subunit vaccine
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