Grossman Z, Winocour E, Berns K I
Virology. 1984 Apr 15;134(1):125-37. doi: 10.1016/0042-6822(84)90278-2.
Recombination between simian virus 40 (SV40) and adeno-associated virus (AAV) has been detected, by infectious center in situ plaque hybridization procedures, after both DNA contransfection and virion coinfection of monkey BSC-1 cells. The number of cells producing recombinants (1 in a 1000) was the same irrespective of the way in which the SV40 and AAV genomes were delivered to the cell, despite the fact that 5-10 times more cells were infected after virion coinfection. Several other dosage-response parameters of the recombination process consequent to virion coinfection were comparable to those after DNA cotransfection. The sole difference observed between the two infection systems was that the SV40/AAV recombinants formed after virion coinfection contained an inordinately high proportion of AAV terminal DNA sequences. By separating the SV40 and AAV infections in time, such that the AAV infection was delayed until after certain events in the SV40 cycle had taken place, an optimum phase for recombination in the SV40 cycle was identified. This phase occurs a few hours after infection, well before the onset of SV40 DNA replication and the synthesis of SV40-specific early proteins.
通过感染中心原位菌斑杂交程序,在猴BSC-1细胞的DNA共转染和病毒粒子共感染后,已检测到猿猴病毒40(SV40)和腺相关病毒(AAV)之间的重组。产生重组体的细胞数量(千分之一)是相同的,无论SV40和AAV基因组递送至细胞的方式如何,尽管在病毒粒子共感染后被感染的细胞多5至10倍。病毒粒子共感染后重组过程的其他几个剂量反应参数与DNA共转染后的参数相当。在两种感染系统之间观察到的唯一差异是,病毒粒子共感染后形成的SV40/AAV重组体含有异常高比例的AAV末端DNA序列。通过及时分离SV40和AAV感染,使AAV感染延迟至SV40周期中的某些事件发生之后,确定了SV40周期中重组的最佳阶段。该阶段在感染后数小时出现,远在SV40 DNA复制开始和SV40特异性早期蛋白合成之前。