ROTAVIRUS-LIKE PARTICLE PRODUCTION IN PLANTS

A method of producing a virus-like particle (VLP) in a plant is provided. The method comprises introducing a first nucleic acid into the plant， or portion of the plant. The first nucleic acid comprising a first regulatory region active in the plant operatively linked to a nucleotide sequence encoding one or more rotavirus structural protein for example but not limited to rotavirus protein VP2. The nucleotide sequence may further comprise one or more than one amplification element and/or a compartment targeting sequence. A second nucleic acid might be introduced into the plant， or portion of the plant. The second nucleic acid comprises a second regulatory region active in the plant and operatively linked to a nucleotide sequence encoding one or more rotavirus structural protein， for example but not limited to rotavirus protein VP6. Optionally， a third nucleic acid and/ or fourth nucleic acid might be introduced into the plant， or portion of the plant. The third nucleic acid comprises a third regulatory region active in the plant and operatively linked to a nucleotide sequence encoding one or more rotavirus structural protein， for example but not limited to rotavirus protein VP4. The fourth nucleic acid comprises a fourth regulatory region active in the plant and operatively linked to a nucleotide sequence encoding one or more rotavirus structural protein， for example but not limited to rotavirus protein VP7. The plant or portion of the plant is incubated under conditions that permit the expression of the nucleic acids， thereby producing the VLP.

本发明公开了一种在植物中制备类病毒颗粒（virus-like particle, VLP）的方法。该方法包括将第一核酸导入该植物或其部分组织中。所述第一核酸包含在植物中具有活性的第一调控区域，其与编码一种或多种轮状病毒结构蛋白的核苷酸序列可操作地连接，所述轮状病毒结构蛋白示例性而非限制性地包括VP2蛋白。该核苷酸序列还可包含一个或多个扩增元件以及/或者区室靶向序列。可任选地向该植物或其部分组织中导入第二核酸，所述第二核酸包含在植物中具有活性的第二调控区域，其与编码一种或多种轮状病毒结构蛋白的核苷酸序列可操作地连接，所述轮状病毒结构蛋白示例性而非限制性地包括VP6蛋白。可选地，还可向该植物或其部分组织中导入第三核酸和/或第四核酸：所述第三核酸包含在植物中具有活性的第三调控区域，其与编码一种或多种轮状病毒结构蛋白的核苷酸序列可操作地连接，所述轮状病毒结构蛋白示例性而非限制性地包括VP4蛋白；所述第四核酸包含在植物中具有活性的第四调控区域，其与编码一种或多种轮状病毒结构蛋白的核苷酸序列可操作地连接，所述轮状病毒结构蛋白示例性而非限制性地包括VP7蛋白。将该植物或其部分组织置于允许核酸表达的条件下培养，即可获得所述类病毒颗粒。