Swoveland P T
J Virol Methods. 1986 Jul;13(4):333-41. doi: 10.1016/0166-0934(86)90058-3.
A method has been developed to isolate measles virus proteins from infected hamster brain tissue. Suckling hamsters inoculated intracerebrally with the HBS strain of measles virus were used in these studies. Viral proteins were isolated from infected brain lysates by affinity chromatography on Sepharose beads coupled with IgG from rabbit hyperimmune anti-measles serum. The eluted proteins were separated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), electrophoretically transferred onto blotting matrix, and immunolabelled with anti-measles antibodies. Individual viral proteins were identified by labelling with monoclonal or monospecific antibodies. All viral proteins except the fusion (F1) protein were identifiable on the immunoblots in relative amounts comparable to purified virions. In addition, a second phosphoprotein (P) band not found in purified virions was present in infected brains and cell cultures infected with HBS or LEC strains of virus. This method should be useful for isolating small quantities of viral proteins from large amounts of tissue, and should make possible the characterization of measles virus proteins in persistently infected CNS tissue.
已开发出一种从感染的仓鼠脑组织中分离麻疹病毒蛋白的方法。在这些研究中，使用了经脑内接种麻疹病毒HBS株的乳仓鼠。通过在与兔超免疫抗麻疹血清的IgG偶联的琼脂糖珠上进行亲和层析，从感染的脑裂解物中分离病毒蛋白。洗脱的蛋白通过SDS-聚丙烯酰胺凝胶电泳（SDS-PAGE）分离，电泳转移到印迹基质上，并用抗麻疹抗体进行免疫标记。通过用单克隆或单特异性抗体标记来鉴定单个病毒蛋白。除融合（F1）蛋白外，所有病毒蛋白在免疫印迹上均能以与纯化病毒粒子相当的相对量被识别。此外，在感染HBS或LEC病毒株的感染脑和细胞培养物中，存在一条在纯化病毒粒子中未发现的第二条磷蛋白（P）带。该方法对于从大量组织中分离少量病毒蛋白应是有用的，并且应使对持续感染的中枢神经系统组织中的麻疹病毒蛋白进行表征成为可能。
