Ng M L, Choo W K, Ho Y L
Department of Microbiology, National University of Singapore, Kent Ridge.
J Virol Methods. 1992 Sep;39(1-2):125-38. doi: 10.1016/0166-0934(92)90131-v.
The types of Kunjin virus-specified proteins present in purified Vero cell plasma membrane were studied. Immunofluorescence of unfixed Kunjin virus-infected whole cell monolayers, indicated that two structural proteins (envelope and prM) and three non-structural proteins (NS1, 3 and 5) were found at the plasma membrane. There was no obvious progressive accumulation of the observed antigens over the time periods between 8 to 24 h p.i. Thus SDS-PAGE analysis was performed using purified radiolabelled Vero cell plasma membranes. From the protein profiles, all five antigens detected by immunofluorescent staining were also present. In addition, two smaller molecular weight non-structural proteins NS4B and NS2B were also observed. Generally, all the non-structural proteins found in the purified plasma membranes were of the same molecular weights as those found in infected whole cell lysate. Interestingly, both the structural proteins, i.e., envelope (E) and prM proteins in the plasma membrane sample were of higher molecular weights as compared to the counterparts in the infected whole cell lysate. The envelope protein of purified extracellular Kunjin virus was also lower in molecular weight compared to the same protein in the plasma membrane.
对纯化的Vero细胞质膜中存在的库京病毒特异性蛋白类型进行了研究。未固定的库京病毒感染的全细胞单层的免疫荧光显示，在质膜中发现了两种结构蛋白（包膜蛋白和prM蛋白）和三种非结构蛋白（NS1、3和5）。在感染后8至24小时的时间段内，观察到的抗原没有明显的逐渐积累。因此，使用纯化的放射性标记的Vero细胞质膜进行了SDS-PAGE分析。从蛋白质谱中可以看出，免疫荧光染色检测到的所有五种抗原也都存在。此外，还观察到了两种分子量较小的非结构蛋白NS4B和NS2B。一般来说，在纯化的质膜中发现的所有非结构蛋白的分子量与在感染的全细胞裂解物中发现的分子量相同。有趣的是，与感染的全细胞裂解物中的对应物相比，质膜样品中的两种结构蛋白，即包膜（E）蛋白和prM蛋白的分子量更高。纯化的细胞外库京病毒的包膜蛋白的分子量也比质膜中的相同蛋白低。
