Matsumura T, Itchoda N, Tsunemitsu H
National Institute of Advanced Industrial and Science Technology, Sapporo, Japan.
Arch Virol. 2002 Jun;147(6):1263-70. doi: 10.1007/s00705-002-0808-4.
We report here the production of transgenic potato plants expressing the major capsid protein VP6 of bovine group A rotavirus (GAR). Transgenic plants under the control of a cauliflower mosaic virus 35S promoter, or a modified promoter linked to the tobacco mosaic virus 5'-untranslated sequence were positive for GAR antigens by ELISA. The expressed protein was consistent in size with VP6 of GAR by Western blot assay. The presence of the VP6 gene and its transcript was detected by PCR and RT-PCR. Adult BALB/c mice were immunized intraperitoneally with concentrated transgenic potato extracts emulsified in Freund's adjuvant. Sera collected after immunization showed the anti-VP6 response in ELISA and Western blot assay. These results suggest that the immunogenic VP6 protein expressed in plants could be useful for the preparation of diagnostic reagents.
我们在此报告表达牛A组轮状病毒（GAR）主要衣壳蛋白VP6的转基因马铃薯植株的培育情况。受花椰菜花叶病毒35S启动子或与烟草花叶病毒5'-非翻译序列相连的修饰启动子控制的转基因植株，通过ELISA检测GAR抗原呈阳性。通过蛋白质印迹分析，所表达的蛋白在大小上与GAR的VP6一致。通过PCR和RT-PCR检测到VP6基因及其转录本的存在。成年BALB/c小鼠用弗氏佐剂乳化的浓缩转基因马铃薯提取物进行腹腔免疫。免疫后收集的血清在ELISA和蛋白质印迹分析中显示出抗VP6反应。这些结果表明，在植物中表达的具有免疫原性的VP6蛋白可用于制备诊断试剂。
