Induction of a virus-specific antibody response to foot and mouth disease virus using the structural protein VP1 expressed in transgenic potato plants
Induction of a virus-specific antibody response to foot and mouth disease virus using the structural protein VP1 expressed in transgenic potato plants
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DOI：
10.1089/08828240151061383
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发表时间：
2001-01-01
期刊：
影响因子：
2.2
通讯作者：
Borca, MV
中科院分区：
文献类型：
--
作者：
Carrillo, C;Wigdorovitz, A;Borca, MV
我们最近在不同的转基因植物中表达了口蹄疫病毒(FMDV)结构蛋白VP1的口服和亲本免疫原性。这些结果清楚地表明，有必要增加外源基因在转基因植物中的表达，以避免提纯和/或浓缩目标抗原的额外步骤。在这里，我们报道了在S35花椰菜花叶病毒(CaMV 35S)启动子的单拷贝(PRok2)或双拷贝(PRok3)调控活性下克隆的含有VP1基因的转基因马铃薯植株的生产，作为提高VP1基因表达水平的策略。用聚合酶链式反应(PCR)和逆转录聚合酶链式反应(RT-PCR)证实了VP1基因在植物中的存在。结果表明，尽管免疫动物产生了口蹄疫病毒VP1的特异性抗体反应和对实验攻击的保护，但从pRok2或pRok3转化植物获得的植物提取物的免疫活性没有显著差异，这些结果证实了以前利用其他植物作为抗原表达载体的结果，并表明至少在马铃薯系统中，使用双CaMV 35S启动子并不能显著提高VP1的表达水平。
We have recently communicated the oral and parental immunogenicity of the structural protein VP1 of foot and mouth disease virus (FMDV) expressed in different transgenic plants. Those results clearly indicated the necessity of increasing the expression of the foreign genes in the transgenic plant to avoid additional steps toward the purification and/or concentration of the antigen of interest. Here, we report the production of transgenic potatoes plants containing the VP1 gene cloned under the regulatory activity of either a single (pRok2) or a double (pRok3) copy of the S35 cauliflower mosaic virus (CaMV 35S) promoter, as a strategy for increasing the level of VP1 gene expression. The presence of the VP1 gene in the plants was confirmed by polymerase chain reaction (PCR) and its specific transcription activity was demonstrated by reverse transcriptase-polymerase chain reaction (RT-PCR), The results showed that, although the immunized animals presented a FMDV VP1 specific antibody response and protection against the experimental challenge, no significant differences were demonstrated in the immunizing activity of plant extracts obtained from the pRok2 or pRok3 transformed plants, These results confirm those previously obtained using other plant species allowing the possibility of using plants as antigen expression vectors, and demonstrated that at least in the potato system, the use of double CaMV 35S promoter does not cause a significant increase in the level of the VP1 expressed.