Spandidos D A, Graham A F
J Virol. 1976 Apr;18(1):117-23. doi: 10.1128/JVI.18.1.117-123.1976.
In standard pairwise crosses there was no detectable recombination between defective reovirus lacking the largest genomic segment and prototypes of the seven known classes of ts mutants. However, in such crosses between R2A (201) and the various prototypes frequencies of ts+ recombinants between 2.6 and 6.1% were observed, as others have found (Fields, 1971; Fields and Joklik, 1969). An infectious center assay was devised to measure recombination in this system, and it was found that all mixedly infected cells gave rise to ts+ recombinants in crosses between prototype ts mutants, but no recombination was detectable when the defective virus was crossed with three different ts mutants. The ts mutation of mutant R2A (201) was efficiently rescued when crossed with UV-inactivated wild-type virus but not when crossed with UV-inactivated defective virus. It is concluded from these various experiments that if there is any recombination between these defective reovirions and any known class of ts mutants it is too low to be measured by methods presently available. The kinetics of recombination were measured in cells mixedly infected with R2A (201) and R2B (352) mutants. At the earliest time progeny virus could be found in the cells the frequency of ts+ recombinants was 4.5%, and this frequency remained unchanged despite a subsequent 1,000-fold increase in progeny virus.
在标准的成对杂交中,缺乏最大基因组片段的缺陷呼肠孤病毒与七种已知类型的温度敏感(ts)突变体的原型之间未检测到重组。然而,在R2A(201)与各种原型之间的此类杂交中,观察到ts +重组体的频率在2.6%至6.1%之间,正如其他人所发现的那样(菲尔兹,1971年;菲尔兹和乔克利克,1969年)。设计了一种感染中心测定法来测量该系统中的重组,结果发现,在原型ts突变体之间的杂交中,所有混合感染的细胞都产生了ts +重组体,但当缺陷病毒与三种不同的ts突变体杂交时,未检测到重组。突变体R2A(201)的ts突变在与紫外线灭活的野生型病毒杂交时能有效拯救,但与紫外线灭活的缺陷病毒杂交时则不能。从这些各种实验得出的结论是,如果这些缺陷呼肠孤病毒与任何已知类型的ts突变体之间存在任何重组,其水平也太低,无法用目前可用的方法测量。在混合感染R2A(201)和R2B(352)突变体的细胞中测量了重组动力学。在最早能在细胞中发现子代病毒的时候,ts +重组体的频率为4.5%,尽管子代病毒随后增加了1000倍,但该频率仍保持不变。