Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice
Cholera toxin B protein in transgenic tomato fruit induces systemic immune response in mice
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DOI：
10.1007/s11248-006-9023-5
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发表时间：
2007-04-01
影响因子：
3
通讯作者：
Yu, Shou-Yi
中科院分区：
文献类型：
--
作者：
Jiang, Xiao-Ling;He, Zhu-Mei;Yu, Shou-Yi
霍乱毒素B（CT B）亚单位是一种特性良好的霍乱抗原。转基因植物可以提供廉价、安全的可食用CTB疫苗来源，可能是生产植物疫苗的最佳候选者之一。本研究旨在利用农杆菌介导的方法，在番茄果实特异性E8启动子的调控下，获得转CTB基因番茄，尤其是在成熟番茄果实中表达CTB蛋白的转基因番茄。利用PCR和Southern印迹分析筛选出转基因植株。从转基因植株的叶、茎和果实组织中提取的外源蛋白经ELISA和Western blot检测表明，CTB蛋白在成熟果实中特异表达，最高表达量为可溶性总蛋白的0.081%。用成熟的转基因番茄果实灌胃小鼠诱导血清和粘膜CTB特异性抗体。这些结果证明了CTB蛋白在转基因番茄中的免疫原性，并为探索CTB在番茄霍乱可食疫苗开发中的应用提供了相当的基础。转基因果实中rCTB抗原导致的抗体滴度比等量的外源CTB低得多，表明果实的纤维组织对外源CTB蛋白在小鼠消化道中的蛋白酶降解的保护作用。
Cholera toxin B (CTB) subunit is a well-characterized antigen against cholera. Transgenic plants can offer an inexpensive and safe source of edible CTB vaccine and may be one of the best candidates for the production of plant vaccines. The present study aimed to develop transgenic tomato expressing CTB protein, especially in the ripening tomato fruit under the control of the tomato fruit-specific E8 promoter by using Agrobacterium-mediated transformation. Transgenic plants were selected using PCR and Southern blot analysis. Exogenous protein extracted from leaf, stem, and fruit tissues of transgenic plants was detected by ELISA and Western blot analysis, showing specific expression in the ripening fruit, with the highest amount of CTB protein being 0.081% of total soluble protein. Gavage of mice with ripe transgenic tomato fruits induced both serum and mucosal CTB specific antibodies. These results demonstrate the immunogenicity of the CTB protein in transgenic tomato and provide a considerable basis for exploring the utilization of CTB in the development of tomato-based edible vaccine against cholera. The rCTB antigen resulted in much lower antibody titers than an equal amount of exgenous CTB in trangenic fruits, suggesting the protective effect of the fibrous tissue of the fruit to the exogenous CTB protein against the degradation of protease in the digestive tracts of mice.