Carrascosa A L, del Val M, Santarén J F, Viñuela E
J Virol. 1985 May;54(2):337-44. doi: 10.1128/JVI.54.2.337-344.1985.
We describe a method for African swine fever (ASF) virus purification based on equilibrium centrifugation in Percoll density gradients of extracellular virions produced in infected VERO cells that yielded about 15 +/- 9% recovery of the starting infectious virus particles. The purified virus preparations were essentially free of a host membrane fraction (vesicles) that could not be separated from the virus by previously described purification methods. The purified virus sedimented as a single component in sucrose velocity gradients with a sedimentation coefficient of 3,500 +/- 300S, showed a DNA-protein ratio of 0.18 +/- 0.02 and a specific infectivity of 2.7 X 10(7) PFU/micrograms of protein, and remained fully infectious after storage at -70 degrees C for at least 7 months. The relative molecular weights of the 34 polypeptides detected in purified virus particles ranged from 10,000 to 150,000. Some of these proteins were probably cellular components that might account for the reactivity of purified virus with antiserum against VERO cells.
我们描述了一种非洲猪瘟(ASF)病毒纯化方法,该方法基于在感染的VERO细胞中产生的细胞外病毒粒子的Percoll密度梯度平衡离心,起始感染性病毒粒子的回收率约为15±9%。纯化的病毒制剂基本不含宿主膜成分(囊泡),而这些成分用先前描述的纯化方法无法与病毒分离。纯化的病毒在蔗糖速度梯度中作为单一成分沉降,沉降系数为3500±300S,DNA与蛋白质的比率为0.18±0.02,比感染性为2.7×10⁷ PFU/微克蛋白质,并且在-70℃下储存至少7个月后仍保持完全感染性。在纯化的病毒粒子中检测到的34种多肽的相对分子量范围为10,000至150,000。其中一些蛋白质可能是细胞成分,这可能解释了纯化病毒与抗VERO细胞抗血清的反应性。