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    • 5. 发明授权
    • Methods for rescue of RNA viruses
    • 拯救RNA病毒的方法
    • US06673572B2
    • 2004-01-06
    • US10261961
    • 2002-10-01
    • Christopher L. ParksMohinderjit S. SidhuStephen A. UdemGerald R. Kovacs
    • Christopher L. ParksMohinderjit S. SidhuStephen A. UdemGerald R. Kovacs
    • C12P2106
    • C07K14/005A61K2039/525C12N7/00C12N2760/18422C12N2760/18451C12N2760/18622C12N2760/18651
    • This invention relates to improved methods for producing nonsegmented, negative-sense, single-stranded RNA viruses of the Order designated Mononegavirales virus, including embodiments relating to methods of producing such viruses as attenuated and/or infectious viruses, such as Measles virus (MV) and respiratory syncytial virus (RSV). One method for producing a recombinant virus from the Order Mononegavirales comprises (a) in at least one host cell, conducting transfection of a rescue composition which comprises (i) a transcription vector comprising an isolated nucleic acid molecule which comprises polynucleotide sequence encoding a genome or antigenome of a nonsegmented, negative-sense, single stranded RNA virus of the Order Mononegavirales and (ii) at least one expression vector which comprises at least one isolated nucleic acid molecule encoding the trans-acting proteins necessary for encapsidation, transcription and replication; in a host cell under conditions sufficient to permit the co-expression of these vectors and the production of the recombinant virus; (b) heating the transfected rescue composition to an effective heat shock temperature under conditions sufficient to increase the recovery of the recombinant virus; and optionally, (c) harvesting the resulting recombinant virus.
    • 本发明涉及用于产生命名为单细胞病毒(Mononegavirales)病毒的非分段,负义的单链RNA病毒的改进方法,其包括与诸如麻疹病毒(MV)等减毒和/或感染性病毒一起生产此类病毒的方法, 和呼吸道合胞病毒(RSV)。 从单顺序单克隆抗体制备重组病毒的一种方法包括(a)在至少一个宿主细胞中,进行拯救组合物的转染,其包含(i)包含分离的核酸分子的转录载体,其包含编码基因组或 单核苷酸单克隆的非分段的,负义的单链RNA病毒的抗原组成单元和(ii)至少一种表达载体,其包含至少一个分离的核酸分子,所述核酸分子编码壳化,转录和复制所必需的反式作用蛋白; 在足以允许这些载体的共表达和重组病毒的产生的条件下在宿主细胞中; (b)在足以增加重组病毒的恢复的条件下将转染的救援组合物加热至有效的热休克温度; 和(c)收获所得的重组病毒。