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    • 24. 发明公开
    • cDNA로부터의 비분절 네가티브 쇄 RNA 바이러스의회수를 위한 개선된 방법
    • 用于从CDNA恢复非分离的负面条件的病毒RNA病毒的改进方法
    • KR1020060028691A
    • 2006-03-31
    • KR1020057023762
    • 2004-06-08
    • 와이어쓰 엘엘씨
    • 팍스크리스토퍼엘우뎀스티븐에이시두모딘데르지트에스
    • C12N15/86C12N15/10C12N7/00
    • C07K14/005A61K39/00A61K48/00A61K2039/5254A61K2039/5256A61K2039/5258C12N7/00C12N15/86C12N2760/18022C12N2760/18422C12N2760/18451C12N2760/18452C12N2760/18551C12N2760/18552C12N2760/18622C12N2760/18651C12N2760/18652C12N2760/18662C12N2760/20243C12N2760/20251C12N2760/20252C12N2760/20262
    • Methods for producing infectious, non-segmented, negative-stranded RNA viruses of the Order Mononegavirales are provided that involve coexpression of a viral cDNA along with essential viral proteins, N, P, and L in a host cell transiently transfected with an expression vector encoding an RNA polymerase. In alternate methods, after the host cell is transfected with a viral cDNA expression vector and one or more vectors encoding the RNA polymerase, N protein, P protein, and L protein, the host cell is exposed to an effective heat shock under conditions sufficient to increase recovery of the recombinant virus. In other alternate embodiments, the host cells are transferred after viral rescue begins into co-culture with a plaque expansion cell, typically a monolayer of expansion cells, and the assembled infectious, non-segmented, negative-stranded RNA virus is recovered from the co-culture. Also provided within the invention are compositions for producing infectious, non-segmented, negative-stranded RNA virus of the Order Mononegavirales, recombinant viruses produced using the foregoing methods and compositions, and immunogenic compositions and methods employing the recombinant viruses. In additional embodiments, the methods and compositions of the invention are employed to produce growth- or replication-defective non-segmented negative-stranded RNA viruses and subviral particles.
    • 提供了产生单宁单胞菌属的感染性,非分段的负链RNA病毒的方法,其涉及病毒cDNA与基本病毒蛋白N,P和L共表达于用表达载体编码瞬时转染的宿主细胞中 RNA聚合酶。 在替代方法中,在用病毒cDNA表达载体和编码RNA聚合酶,N蛋白,P蛋白和L蛋白的一种或多种载体转染宿主细胞后,将宿主细胞暴露于有效的热休克, 增加重组病毒的恢复。 在其它替代实施方案中,宿主细胞在病毒救治开始与噬菌斑扩增细胞(通常为单层扩增细胞)共培养之后被转移,并且从共同体中回收组装的感染性,非分段的负链RNA病毒 -文化。 在本发明中还提供了用于产生单宁属(Mononegavirales)的感染性,非分段的负链RNA病毒的组合物,使用前述方法和组合物产生的重组病毒,以及使用重组病毒的免疫原性组合物和方法。 在另外的实施方案中,本发明的方法和组合物用于产生生长或复制缺陷型非分段负链RNA病毒和亚病毒颗粒。