The present invention relates to recombinant Adenoviruses (Ad) which function as helper viruses to gutted Ad viruses lacking viral coding sequences. The recombinant Ad of the present invention comprise a binding site of the
E. coli
Lac repressor protein embedded within the packaging domain. Available Lac repressor protein binds to its operator site within the helper virus packaging domain, precluding the binding of natural packaging factors. The present invention also provides recombinant Ad which in addition to having a binding site for the Lac repressor protein within the packaging domain, also comprise coding sequence for the Lac repressor protein under the control of a promoter which functions in producer cells. Methods for suppressing packaging of helper Ad during packaging of a gutted Ad vector are also provided as are methods of producing recombinant helper Ad using lactose or lactose derivative as a regulatory molecule. Temperature shift may also be used to regulate growth of a recombinant helper Ad. In addition, methods of producing a gutted Ad vector substantially free of helper Ad are provided.
本发明涉及
重组腺病毒(Ad),
重组腺病毒可作为缺乏病毒编码序列的内脏腺病毒的辅助病毒。本发明的
重组 Ad 包括一个与
大肠杆菌
Lac 抑制蛋白的结合位点。可用的 Lac 抑制蛋白与辅助病毒包装结构域内的操作者位点结合,排除了天然包装因子的结合。本发明还提供了
重组 Ad,这种 Ad 除了在包装结构域内具有 Lac 抑制蛋白的结合位点外,还包括 Lac 抑制蛋白的编码序列,该编码序列受在生产者细胞中起作用的启动子控制。此外,还提供了在包装去内脏 Ad 载体期间抑制辅助 Ad 包装的方法,以及使用
乳糖或
乳糖衍
生物作为调节分子生产
重组辅助 Ad 的方法。温度变化也可用于调节
重组辅助 Ad 的生长。此外,还提供了生产基本上不含辅助性 Ad 的开胃 Ad 载体的方法。