Narang H K
Ken Bell International, Newcastle-Upon-Tyne, UK.
Res Virol. 1998 Nov-Dec;149(6):375-82. doi: 10.1016/s0923-2516(99)80005-3.
Homogenized brain tissue from scrapie-infected hamsters and uninfected hamsters was subjected to subcellular fractionation to isolate unique tubulofilamentous particles termed "nemaviruses". Nucleic acid was purified from the concentrate by the phenol/chloroform extraction procedure and by alkaline gel electrophoresis; a single band of ssDNA corresponding to about 1.2 kb was visualized. The gel-purified ssDNA was mixed before inoculation with (a) MgCl2, (b) lipofectin, (c) ssDNA-binding protein and (d) normal brain homogenate. Hamsters in group b, c and d injected with a mixture of ssDNA developed the clinical disease. Brain pathology revealed generalized vacuolation, while animals injected with ssDNA mixed with MgCl2 and controls remained healthy. These results suggest that ssDNA is the genome of the scrapie agent.
将来自感染瘙痒病的仓鼠和未感染仓鼠的匀浆脑组织进行亚细胞分级分离,以分离出被称为“线虫病毒”的独特管状丝状颗粒。通过苯酚/氯仿提取法和碱性凝胶电泳从浓缩物中纯化核酸;观察到一条对应于约1.2 kb的单链DNA条带。在接种前,将凝胶纯化的单链DNA与(a)氯化镁、(b)脂质体、(c)单链DNA结合蛋白和(d)正常脑匀浆混合。注射单链DNA混合物的b、c和d组仓鼠出现了临床疾病。脑部病理学显示广泛空泡化,而注射了与氯化镁混合的单链DNA的动物和对照组保持健康。这些结果表明单链DNA是瘙痒病病原体的基因组。