Nonoyama M, Watanabe Y, Graham A F
J Virol. 1970 Aug;6(2):226-36. doi: 10.1128/JVI.6.2.226-236.1970.
When purified preparations of stock reovirus, type 3, were digested with chymotrypsin, the virions were converted into two different types of particle. These new particles could be separated from each other by isopycnic centrifugation in cesium chloride gradients. One particle banded at a buoyant density of 1.43 g/cm(3), the other at a density of 1.415 g/cm(3). The former particle is termed the heavy (H) particle, the latter is the light (L) particle. The ratio of H/L particles varied between 0.5 and 0.25 in various purified preparations of virus. In electron micrographs, both H and L particles had the appearance and dimensions of viral cores. H particles were infectious for L cells. When plaques formed by stock virus, or by H particles, were picked and propagated in L cells, the majority of the clones gave rise only to H particles on chymotrypsin digestion. On continued serial passage of the clones, virions containing L particles again appeared in the progeny. The simplest explanation of these results was that stock virus was comprised of two populations of virions. One type of virion which contained H particles was infectious, whereas the other, which contained L particles, was not itself infectious and could replicate only in cells coinfected with an H particle virion. Added weight was given to this hypothesis by two observations. First, a small but definite separation of H and L virions could be achieved by isopycnic centrifugation in a gradient of cesium chloride. Second, L particles and virions containing L particles were both shown to lack the largest of the ten segments of double-stranded ribonucleic acid genome. Thus, L particle virions have defective genomes.
当用胰凝乳蛋白酶消化纯化的3型呼肠孤病毒储备制剂时,病毒粒子被转化为两种不同类型的颗粒。这些新颗粒可以通过在氯化铯梯度中进行等密度离心彼此分离。一种颗粒在浮力密度为1.43 g/cm³处形成条带,另一种在密度为1.415 g/cm³处形成条带。前一种颗粒称为重(H)颗粒,后一种是轻(L)颗粒。在各种纯化的病毒制剂中,H/L颗粒的比例在0.5至0.25之间变化。在电子显微镜照片中,H颗粒和L颗粒都具有病毒核心的外观和尺寸。H颗粒对L细胞具有感染性。当由储备病毒或H颗粒形成的噬斑被挑选出来并在L细胞中传代培养时,大多数克隆在胰凝乳蛋白酶消化后只产生H颗粒。随着克隆的持续传代,含有L颗粒的病毒粒子再次出现在子代中。对这些结果最简单的解释是储备病毒由两种病毒粒子群体组成。一种含有H颗粒的病毒粒子具有感染性,而另一种含有L颗粒的病毒粒子本身没有感染性,只能在与H颗粒病毒粒子共感染的细胞中复制。两项观察结果支持了这一假设。第一,通过在氯化铯梯度中进行等密度离心,可以实现H和L病毒粒子的小而明确的分离。第二,L颗粒和含有L颗粒的病毒粒子都被证明缺乏双链核糖核酸基因组的十个片段中最大的那个片段。因此,L颗粒病毒粒子具有缺陷基因组。