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烟草种子中高浓度的禽类(H5N1)流感血凝素抗原。

High accumulation in tobacco seeds of hemagglutinin antigen from avian (H5N1) influenza.

机构信息

Plant Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), PO Box 6162, 10600, Havana, Havana, Cuba.

Department of Carbohydrate Chemistry, Center for Genetic Engineering and Biotechnology (CIGB), Havana, Cuba.

出版信息

Transgenic Res. 2017 Dec;26(6):775-789. doi: 10.1007/s11248-017-0047-9. Epub 2017 Oct 6.

Abstract

Tobacco seeds can be used as a cost effective system for production of recombinant vaccines. Avian influenza is an important respiratory pathogen that causes a high degree of mortality and becomes a serious threat for the poultry industry. A safe vaccine against avian flu produced at low cost could help to prevent future outbreaks. We have genetically engineered tobacco plants to express extracellular domain of hemagglutinin protein from H5N1 avian influenza virus as an inexpensive alternative for production purposes. Two regulatory sequences of seed storage protein genes from Phaseolus vulgaris L. were used to direct the expression, yielding 3.0 mg of the viral antigen per g of seeds. The production and stability of seed-produced recombinant HA protein was characterized by different molecular techniques. The aqueous extract of tobacco seed proteins was used for subcutaneous immunization of chickens, which developed antibodies that inhibited the agglutination of erythrocytes after the second application of the antigen. The feasibility of using tobacco seeds as a vaccine carrier is discussed.

摘要

烟草种子可作为生产重组疫苗的具有成本效益的系统。禽流感是一种重要的呼吸道病原体,可导致高度死亡率,并对家禽业构成严重威胁。低成本生产的针对禽流感的安全疫苗有助于预防未来的爆发。我们已通过基因工程改造烟草植物,以表达 H5N1 禽流感病毒的血凝素蛋白的细胞外结构域,作为生产目的的廉价替代品。来自菜豆的种子贮藏蛋白基因的两个调控序列被用来指导表达,使每克种子产生 3.0 毫克的病毒抗原。通过不同的分子技术对种子产生的重组 HA 蛋白的生产和稳定性进行了表征。烟草种子蛋白的水提物用于鸡的皮下免疫,接种抗原后第二次应用时,鸡产生了可抑制红细胞凝集的抗体。讨论了利用烟草种子作为疫苗载体的可行性。

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