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首页> 外文期刊>Journal of Biotechnology >Expression of accessory colonization factor subunit A (ACFA) of Vibrio cholerae and ACFA fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum)
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Expression of accessory colonization factor subunit A (ACFA) of Vibrio cholerae and ACFA fused to cholera toxin B subunit in transgenic tomato (Solanum lycopersicum)

机译：霍乱弧菌的辅助定殖因子亚基A（ACFA）和与霍乱毒素B亚基融合的ACFA在转基因番茄（Solanum lycopersicum）中的表达

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摘要

In earlier study from our group, cholera toxin B subunit had been expressed in tomato for developing a plant-based vaccine against cholera. In the present investigation, gene for accessory colonization factor (acf) subunit A, earlier reported to be essential for efficient colonization in the intestine, has been expressed in Escherichia coli as well as tomato plants. Gene encoding for a chimeric protein having a fusion of cholera toxin B subunit and accessory colonization factor A was also expressed in tomato to generate more potent combinatorial antigen. CaMV35S promoter with a duplicated enhancer sequence was used for expression of these genes in tomato. Integration of transgenes into tomato genome was confirmed by PCR and Southern hybridization. Expression of the genes was confirmed at transcript and protein levels. Accessory colonization factor A and cholera toxin B subunit fused to this protein accumulated up to 0.25% and 0.08% of total soluble protein, respectively, in the fruits of transgenic plants. Whereas protein purified from E. coli, in combination with cholera toxin B subunit can be used for development of conventional subunit vaccine, tomato fruits expressing these proteins can be used together with tomato plants expressing cholera toxin B subunit for development of oral vaccine against cholera.
机译：在我们小组的早期研究中，霍乱毒素B亚基已在番茄中表达，用于开发基于植物的霍乱疫苗。在本研究中，先前报道在肠道中有效定殖必不可少的辅助定殖因子（acf）亚基A的基因已在大肠杆菌和番茄植物中表达。编码具有霍乱毒素B亚基和辅助定殖因子A融合的嵌合蛋白的基因也在番茄中表达以产生更有效的组合抗原。具有重复的增强子序列的CaMV35S启动子用于在番茄中表达这些基因。通过PCR和Southern杂交证实了转基因整合到番茄基因组中。在转录本和蛋白质水平上证实了基因的表达。与该蛋白融合的辅助定居因子A和霍乱毒素B亚基分别在转基因植物的果实中累积高达总可溶性蛋白的0.25％和0.08％。从大肠杆菌中纯化的蛋白质与霍乱毒素B亚基结合可用于常规亚单位疫苗的开发，表达这些蛋白质的番茄果实可与表达霍乱毒素B亚基的番茄植株一起用于开发针对霍乱的口服疫苗。

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