Nadala E C, Loh P C
Department of Microbiology, University of Hawaii, Honolulu 96822, USA.
Dis Aquat Organ. 1998 Jul 30;33(3):231-4. doi: 10.3354/dao033231.
Three separate isolates of white spot virus (WSV) purified from 3 different penaeid shrimp species from different countries were compared morphologically, biochemically, and genomically using the following techniques; negative stain electron microscopy, sodium dodecyl-sulfate polyacrylamide gel electrophoresis/western blot, and restriction fragment length polymorphism (RFLP), respectively. Under the electron microscope, the 3 isolates were indistinguishable. Their nucleoprotein cores exhibited the unique striated structure characteristic of the baculovirus-like agents associated with white spot syndrome. The dimensions of the nucleoprotein cores were also identical for all 3 isolates. SDS-PAGE gels of purified virus preparations showed all 3 to be identical in the position of at least 3 of the most prominent protein bands of WSV, with approximate molecular weights of 19, 23.5, and 27.5 kDa. Western blot analyses also revealed these 3 same protein bands in identical positions for all 3 isolates. RFLP analyses of the viral genomes using Hind III and EcoR 1 enzymes revealed that although the 3 isolates were identical when cut with EcoR I, the isolate from Penaeus japonicus from China was distinguishable from the other 2 genomes (P. monodon from Indonesia and P. setiferus from the U.S.) when cut with Hind III.
从不同国家的3种不同对虾品种中纯化得到的3株白斑病毒(WSV)分离株,分别采用以下技术进行形态学、生物化学和基因组学比较:负染电子显微镜技术、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳/蛋白质免疫印迹法以及限制性片段长度多态性分析(RFLP)。在电子显微镜下,这3株分离株无法区分。它们的核蛋白核心呈现出与白斑综合征相关的杆状病毒样病原体特有的条纹状结构特征。所有3株分离株的核蛋白核心尺寸也相同。纯化病毒制剂的SDS-PAGE凝胶显示,所有3株在WSV至少3条最突出蛋白带的位置上相同,其近似分子量分别为19、23.5和27.5 kDa。蛋白质免疫印迹分析也显示所有3株在相同位置有这3条相同的蛋白带。使用Hind III和EcoR 1酶对病毒基因组进行RFLP分析表明,虽然用EcoR I切割时这3株分离株相同,但用Hind III切割时,来自中国的日本对虾分离株与其他2个基因组(来自印度尼西亚的斑节对虾和来自美国的长缝拟对虾)不同。