Belli B A, Samuel C E
Department of Biological Sciences, University of California, Santa Barbara 93106.
Virology. 1991 Dec;185(2):698-709. doi: 10.1016/0042-6822(91)90541-i.
Polyclonal monospecific antibody was prepared against the reovirus serotype 1 Lang strain nonstructural sigma 1NS protein encoded by the S1 gene. The antibody was serotype-specific. The sigma 1NS protein of reovirus serotype 1, but not reovirus serotype 3, was recognized by the polyclonal antibody in both immunoprecipitation and Western immunoblot assays. The sigma 1NS protein expressed in vector-transfected COS cells was indistinguishable by immunoprecipitation and immunoblot analyses from the authentic sigma 1NS protein synthesized in virus-infected mouse L or monkey COS cells. The temporal appearance of sigma 1NS protein in virus-infected cells was similar to that of the other reovirus proteins. Both sigma 1NS and sigma 1, the two S1 gene products, were observed in the cytoplasm of COS cells by immunofluorescent microscopy, although their staining patterns were distinct from each other. However, sigma 1NS, but not sigma 1 or the other reovirion structural proteins, was also detected in the nucleoli of COS cells. These results suggest that sigma 1NS, like sigma 1, is a serotype-specific reovirus protein, but unlike sigma 1 is localized in part to the cell nucleus.
针对呼肠孤病毒1型朗株由S1基因编码的非结构σ1NS蛋白制备了多克隆单特异性抗体。该抗体具有血清型特异性。在免疫沉淀和Western免疫印迹分析中，呼肠孤病毒1型的σ1NS蛋白能被多克隆抗体识别，而呼肠孤病毒3型的则不能。在载体转染的COS细胞中表达的σ1NS蛋白，通过免疫沉淀和免疫印迹分析，与在病毒感染的小鼠L细胞或猴COS细胞中合成的天然σ1NS蛋白无法区分。病毒感染细胞中σ1NS蛋白的出现时间与其他呼肠孤病毒蛋白相似。通过免疫荧光显微镜观察，在COS细胞的细胞质中同时发现了σ1NS和σ1这两种S1基因产物，尽管它们的染色模式彼此不同。然而，在COS细胞的核仁中也检测到了σ1NS，而未检测到σ1或其他呼肠孤病毒结构蛋白。这些结果表明，σ1NS与σ1一样，是一种血清型特异性的呼肠孤病毒蛋白，但与σ1不同的是，它部分定位于细胞核。
