SEQUENTIAL PROPAGATION OF POLIOVIRUS RNA GENOMES USING IMPROVED TRANSFECTION

使用脊髓灰质炎病毒RNA的转染法已经改进到 纯化的病毒RNA的感染性接近 完整病毒粒子的传染性。使用苯酚提取的RNA与 阳离子洗涤剂脂肪体和悬浮细胞培养，我们设计了 导致细胞100%感染的条件。上一首 转染法的效率降低了10%到100%。如此高水平的 转染法，结合有效的RNA分离程序，提供了一种 用于脊髓灰质炎病毒RNA连续传递的手段，同时绕过 病毒的功能与包膜和去膜有关。因此， 选择压力导致亚基因组复制子的产生 仅保留病毒RNA复制所需的功能。我们是 使用这种方法来遵循自然的选择压力 脊髓灰质炎病毒缺陷干扰基因组在 缺少封装约束。

Transfection procedures using poliovirus RNA have been improved to the point where the infectivity of the purified viral RNA approaches the infectivity of the intact virion. Using phenol extracted RNA with cationic detergent lipsomes and suspension cell cultures, we devised conditions that result in 100% infection of the cells. Previous transfection procedures were 10 to 100 less efficient. This high level of transfection, coupled with efficient RNA isolation procedures, provides a means for the sequential passage of poliovirus RNA while bypassing those viral functions involved in encapsidation and uncoating. Consequently, selective pressures result in the generation of subgenomic replicons retaining only those functions required for viral RNA replication. We are using this approach to follow the natural selective pressures on poliovirus defective interfering genomes when they are passaged in the absence of encapsidation constraints.