Roggendorf M, Pahlke C, Böhm B, Rasshofer R
Max von Pettenkofer Institut for Medical Microbiology and Hygiene, University of Munich, F.R.G.
J Gen Virol. 1987 Nov;68 ( Pt 11):2953-9. doi: 10.1099/0022-1317-68-11-2953.
The number and size of proteins associated with hepatitis delta virus (HDV) from serum and liver (human, chimpanzee and woodchuck) in the acute and chronic stages of HDV infection were analysed by immunoblotting. HDV particles in serum were separated from serum proteins by gel filtration and peak fractions of HDV antigens were subjected to PAGE. Immunoblotting with human anti-HDV-positive sera and 125I-labelled Protein A revealed two bands of 27K and 29K. It was not possible to identify any core-like structure from liver homogenates by CsCl gradient centrifugation. HDV proteins from such gradients were degraded to a size of 14K as determined by immunoblotting. HDV RNA was found in fractions at a density of 1.5 g/ml. However, direct homogenization of liver tissue in gel electrophoresis sample buffer, followed by PAGE and immunoblotting resulted in identification of HDV-associated proteins of 27K and 29K, indicating that HDV proteins in liver tissue are the same size as those in serum, but that they degrade rapidly. There was no difference in size of HDV proteins in liver samples from humans, chimpanzees or woodchucks.
通过免疫印迹法分析了在丁型肝炎病毒（HDV）感染的急性和慢性阶段，来自血清和肝脏（人类、黑猩猩和土拨鼠）的与HDV相关的蛋白质的数量和大小。血清中的HDV颗粒通过凝胶过滤与血清蛋白分离，HDV抗原的峰值组分进行聚丙烯酰胺凝胶电泳（PAGE）。用人抗HDV阳性血清和125I标记的蛋白A进行免疫印迹显示出两条分别为27K和29K的条带。通过氯化铯梯度离心无法从肝脏匀浆中鉴定出任何核心样结构。通过免疫印迹法测定，来自这种梯度的HDV蛋白降解至14K大小。在密度为1.5 g/ml的组分中发现了HDV RNA。然而，将肝脏组织直接在凝胶电泳样品缓冲液中匀浆，随后进行PAGE和免疫印迹，结果鉴定出27K和29K的HDV相关蛋白，表明肝脏组织中的HDV蛋白与血清中的大小相同，但它们降解迅速。来自人类、黑猩猩或土拨鼠的肝脏样品中HDV蛋白的大小没有差异。
