Drzeniek R
Z Naturforsch C Biosci. 1975 Jul-Aug;30(4):523-31. doi: 10.1515/znc-1975-7-818.
Poliovirus is dissociated by urea in the presence of mercaptoethanol or dithiothreitol into RNA and protein(s). Differences exist in the sensitivity of poliovirus type I, II or III toward these agents. The decrease in infectivity titer of the virion is in the range of 10(9). After appropriate dissociation no infectious virions are found in urea-treated poliovirus concentrates. The released viral RNA is infectious. Its infectivity (1 times 10(6) PFU/mug RNA) is comparable to that of phenol extracted RNA. Therefore, urea dissociation is a fast and reliable method for the preparation of infectious poliovirus RNA even at the microliter scale. The dissociation of poliovirus is dependent upon the concentration, temperature and pH. The decrease in virion infectivity is time dependent. It is preceded by a lag phase of different lengths for poliovirus type I, II or III. During the lag phase the virion undergoes configurational changes without losing infectivity. From the observed hyperchromic effect is it obvious that the secondary structure of the RNA inside the virion breaks down, giving rise to expanded forms of poliovirus particles. After the lag phase the RNA is released and the empty capsid dissociated into the appropriate polypeptides.
在巯基乙醇或二硫苏糖醇存在的情况下,尿素可将脊髓灰质炎病毒解离为RNA和蛋白质。脊髓灰质炎病毒I型、II型或III型对这些试剂的敏感性存在差异。病毒粒子的感染性滴度下降幅度在10⁹范围内。经过适当解离后,在经尿素处理的脊髓灰质炎病毒浓缩物中未发现有感染性的病毒粒子。释放出的病毒RNA具有感染性。其感染性(1×10⁶ PFU/μg RNA)与酚提取的RNA相当。因此,即使在微升规模下,尿素解离也是制备感染性脊髓灰质炎病毒RNA的一种快速且可靠的方法。脊髓灰质炎病毒的解离取决于浓度、温度和pH值。病毒粒子感染性的下降是时间依赖性的。脊髓灰质炎病毒I型、II型或III型在下降之前有不同长度的延迟期。在延迟期内,病毒粒子发生构象变化但不丧失感染性。从观察到的增色效应可以明显看出,病毒粒子内部RNA的二级结构被破坏,产生了脊髓灰质炎病毒颗粒的扩展形式。延迟期过后,RNA被释放,空衣壳解离为相应的多肽。