van de Ven W J, Vermorken A J, Onnekink C, Bloemers H P, Bloemendal H
J Virol. 1978 Sep;27(3):595-603. doi: 10.1128/JVI.27.3.595-603.1978.
A preparative method for isolating pure viral envelopes from a type-C RNA tumor virus, Rauscher murine leukemia virus, is described. Fractionation of virions of Rauscher murine leukemia virus was studied after disruption of the virions with the detergents sodium dodecyl sulfate of Nonidet P-40 in combination with ether. Fractionation was performed through flotation in a discontinuous sucrose gradient and, as appeared from electron microscopic examination, a pure viral envelope fraction was obtained in this way. By use of sensitive competition radioimmunoassays or sodium dodecyl sulfate-polyacrylamide gel electrophoresis after immunoprecipitation with polyvalent and monospecific antisera directed against Rauscher murine leukemia virus proteins, the amount of the gag and env gene-encoded structural polypeptides in the virions and the isolated envelope fraction was compared. The predominant viral structural polypeptides in the purified envelope fraction were the env gene-encoded polypeptides gp70, p15(E), and p12(E), whereas, except for p15, there was only a relatively small amount of the gag gene-encoded structural polypeptides in this fraction.
本文描述了一种从C型RNA肿瘤病毒——劳舍尔鼠白血病病毒中分离纯病毒包膜的制备方法。在用十二烷基硫酸钠或Nonidet P-40去污剂结合乙醚破坏劳舍尔鼠白血病病毒粒子后,对其进行分级分离研究。分级分离通过在不连续蔗糖梯度中浮选进行,从电子显微镜检查结果来看,通过这种方式获得了纯病毒包膜级分。使用针对劳舍尔鼠白血病病毒蛋白的多价和单特异性抗血清进行免疫沉淀后,通过灵敏的竞争放射免疫测定法或十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,比较了病毒粒子和分离出的包膜级分中gag和env基因编码的结构多肽的含量。纯化包膜级分中主要的病毒结构多肽是env基因编码的多肽gp70、p15(E)和p12(E),而在该级分中,除了p15外,gag基因编码的结构多肽含量相对较少。