College of Life Science and Technology, Northwest A&F University, Yangling, Shanxi, China.
J Virol Methods. 2012 Apr;181(1):12-7. doi: 10.1016/j.jviromet.2012.01.004. Epub 2012 Jan 16.
Transgenic plants have become developed as bioreactors for producing heterologous proteins and may even form edible vaccines. In the present study, a transgenic rice expressing the capsid precursor polypeptide (P1) gene of foot-and-mouth disease virus (FMDV), under the control of a dual cauliflower mosaic virus (CaMV 35S) promoter, was generated by Agrobacterium-mediated transformation. Southern blot, northern blot, western blot, and ELISA analyses confirmed that the P1 gene was integrated into the transgenic rice and the protein was expressed specifically in the leaves at levels of 0.6-1.3 μg/mg of total soluble protein. After intraperitoneal immunization of mice with crude protein extracts from transgenic rice plants, FMDV-specific neutralizing antibodies were detected. The immunized mice could clear virus from their sera after FMDV challenge. In addition, FMDV-specific mucosal immune responses were detected in mice after oral immunization with protein extracts from transgenic rice plants. Partial virus clearance was obtained after FMDV challenge. These results indicate the potential of using a transgenic rice-based expression system as an alternative bioreactor for FMDV subunit vaccines.
转基因植物已被开发为生产异源蛋白的生物反应器，甚至可能形成可食用的疫苗。本研究通过农杆菌介导的转化方法，生成了一种转基水稻，该水稻表达口蹄疫病毒（FMDV）的衣壳前体多肽（P1）基因，受双花椰菜花叶病毒（CaMV 35S）启动子的控制。Southern blot、northern blot、western blot 和 ELISA 分析证实，P1 基因已整合到转基因水稻中，并且该蛋白特异性地在叶片中表达，总可溶性蛋白水平为 0.6-1.3 μg/mg。用转基因水稻植物的粗蛋白提取物对小鼠进行腹腔免疫后，检测到 FMDV 特异性中和抗体。在 FMDV 攻毒后，免疫小鼠可从血清中清除病毒。此外，用转基因水稻植物的蛋白提取物对小鼠进行口服免疫后，检测到 FMDV 特异性黏膜免疫反应。在 FMDV 攻毒后获得部分病毒清除。这些结果表明，使用基于转基因水稻的表达系统作为 FMDV 亚单位疫苗的替代生物反应器具有潜力。
