Gugerli P
J Virol Methods. 1984 Nov;9(3):249-58. doi: 10.1016/0166-0934(84)90031-4.
Isopycnic centrifugation of plant viruses in density gradients prepared from a new non-ionic medium (Nycodenz) was investigated. Particle density of luteo-, tymo-, nepo-, cocksfoot mild mosaic, tobamo-, hordei-like, potex- and potyviruses in buffered solutions of Nycodenz ranged between 1.23 and 1.28 g/ml and did not strictly reflect their nucleic acid and protein composition. Isopycnic centrifugation of several partially purified viruses yielded preparations which were pure by electron microscopy and, when used as immunogens in rabbit or hen, gave antisera with low titers of antibodies against host plant antigens, as required for enzyme immunoassays. A preparation of purified potato leafroll virus contained a single dominant protein corresponding to the viral coat protein. It is concluded that isopycnic centrifugation in Nycodenz density gradients is particularly useful for the purification and analysis of plant viruses which are unstable in cesium chloride solutions, such as the luteoviruses, and preferable to zonal centrifugation in sucrose density gradients for the purification of rod-shaped multipartite viruses or viruses which aggregate.
研究了在由一种新型非离子介质（Nycodenz）制备的密度梯度中对植物病毒进行等密度离心的方法。在Nycodenz缓冲溶液中，黄症病毒属、烟草花叶病毒属、线虫传多面体病毒属、鸭茅轻型花叶病毒、烟草花叶病毒属、大麦病毒样、马铃薯X病毒属和马铃薯Y病毒属的粒子密度在1.23至1.28克/毫升之间，并未严格反映其核酸和蛋白质组成。对几种部分纯化的病毒进行等密度离心，得到的制剂通过电子显微镜观察是纯的，并且当用作兔或鸡的免疫原时，能产生针对宿主植物抗原的抗体滴度较低的抗血清，这是酶免疫测定所需要的。纯化的马铃薯卷叶病毒制剂含有一种单一的主要蛋白质，与病毒外壳蛋白相对应。得出的结论是，在Nycodenz密度梯度中进行等密度离心对于纯化和分析在氯化铯溶液中不稳定的植物病毒（如黄症病毒）特别有用，并且对于纯化杆状多分体病毒或聚集的病毒而言，比在蔗糖密度梯度中进行区带离心更可取。
