Abstract：
In this protocol, we outline how to produce a chimeric viral vaccine in a biosafety level 1 (BSL1) environment. An animal viral vector RNA encapsidated with tobacco mosaic virus (TMV) coat protein can be fully assembled in planta. Agrobacterium cultures containing each component are inoculated together into tobacco leaves and the self-assembled hybrid chimeric viral vaccine is harvested 4 days later and purified with a simple PEG precipitation. The viral RNA delivery vector is derived from the BSL1 insect virus, Flock House virus (FHV), and replicates in human and animal cells but does not spread systemically. A polyethylene glycol purification protocol is also provided to collect and purify these vaccines for immunological tests. In this update, we also provide a protocol for in trans co-inoculation of a modified FHV protein A, which significantly increased the yield of in planta chimeric viral vaccine.
摘要：
在这个协议中,我们概述了如何在生物安全1级(BSL1)环境中生产嵌合病毒疫苗.用烟草花叶病毒（TMV）外壳蛋白包裹的动物病毒载体RNA可以在植物中完全组装。将含有每种组分的农杆菌培养物一起接种到烟草叶中,4天后收获自组装的杂合嵌合病毒疫苗,并用简单的PEG沉淀纯化。病毒RNA递送载体来源于BSL1昆虫病毒，羊室病毒(FHV),并在人和动物细胞中复制，但不会全身传播。还提供聚乙二醇纯化方案以收集和纯化这些疫苗用于免疫学测试。在此更新中，我们还提供了一种用于反式共接种修饰的FHV蛋白A的方案，这显著提高了植物嵌合病毒疫苗的产量。
