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COMPOSITIONS AND METHODS USEFUL FOR PRODUCTION OF RECOMBINANT VIRUSES WHICH REQUIRE HELPER VIRUSES

机译:用于生产需要辅助病毒的重组病毒的组合物和方法

摘要

A method of producing recombinant viruses which require helper viruses forpackaging is provided. The method uses a recombinant helper virus which hasbeen designed to contain at least one rare-cutting restriction site (e.g., forI-SceI), a viral vector which is helper-dependent for packaging, and arecombinant host cell which is capable of expressing a rare-cuttingrestriction enzyme (e.g., I-SceI). The method involves transfecting orinfecting the host cell with the helper virus and viral vector and incubatingthe cell under conditions which permit encapsidation of the viral vector in anadenovirus capside. Thereafter, the I-SceI expressed by the host cell digeststhe helper virus, permitting ready separation of the digested fragments of thehelper virus from the packaged recombinant virus.
机译:一种产生重组病毒的方法,该重组病毒需要辅助病毒用于提供包装。该方法使用重组辅助病毒被设计为包含至少一个罕见的限制性酶切位点(例如I-SceI),包装依赖于助手的病毒载体和表达罕见切割的重组宿主细胞限制性内切酶(例如I-SceI)。该方法涉及转染或用辅助病毒和病毒载体感染宿主细胞并孵育在允许病毒载体衣壳化的条件下腺病毒。此后,宿主细胞表达的I-SceI消化辅助病毒,可以迅速分离出消化后的片段包装的重组病毒中的辅助病毒。

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