Purification of Rabies Virus Grown in Tissue Culture
Purification of Rabies Virus Grown in Tissue Culture
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组织培养中生长的狂犬病病毒的纯化
DOI：
10.1128/jvi.2.8.836-849.1968
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发表时间：
1968
影响因子：
5.4
通讯作者：
H. Koprowski
中科院分区：
文献类型：
--
作者：
F. Sokol;E. Kuwert;T. Wiktor;K. Hummeler;H. Koprowski
在补充有牛血清白蛋白的培养基存在下，在 BHK21 细胞的单层培养物中生长的胞外狂犬病病毒通过以下程序纯化。用乙酸锌从感染性组织培养液中沉淀病毒，并将其重悬于乙二胺四乙酸溶液中。将悬浮液通过Sephadex柱过滤并用核糖核酸酶和脱氧核糖核酸酶处理。然后通过高速离心沉淀病毒颗粒并重悬于缓冲溶液中。通过在蔗糖密度梯度中离心对病毒进行显带是纯化过程的最后一步。纯化的制剂含有不同长度的子弹形病毒颗粒和少量（最多 5%）的宿主细胞污染物质。大多数病毒颗粒是“完整的”，即 180 nm 长，但一些病毒颗粒更短。病毒体的长度分布是非随机的。较短的病毒体似乎不具有感染性，并且血凝活性显着降低。病毒粒子的补体固定活性和核糖核酸与蛋白质的比率与病毒颗粒的长度无关。尽管胞外病毒和胞内病毒的特性相似，但该方法不适用于胞内狂犬病病毒的纯化。
Extracellular rabies virus, grown in monolayer cultures of BHK21 cells in the presence of medium supplemented with bovine serum albumin, was purified by the following procedure. Virus was precipitated from infectious tissue culture fluid by zinc acetate and was resuspended in a solution of ethylenediaminetetraacetate. The suspension was filtered through a Sephadex column and was treated with ribonuclease and deoxyribonuclease. The virions were then pelleted by centrifugation at high speed and were resuspended in buffer solution. Banding of the virus by centrifugation in a sucrose density gradient was the final step in the purification procedure. Purified preparations contained bullet-shaped virus particles of variable length and little (up to 5%) contaminating host-cell material. Most of the virions were “complete”, i.e., 180 nm long, but some virus particles were shorter. The length distribution of the virions was nonrandom. Shorter virions seemed to be noninfectious and showed markedly decreased hemagglutinating activity. The complement-fixing activity and the ribonucleic acid to protein ratio of the virions were not related to the length of the virus particles. Although the properties of extracellular and intracellular viruses were similar, the procedure was not suitable for purification of intracellular rabies virus.