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Comparison of Genome Size and Synthesis of Structural Proteins of Hirame Rhabdovirus, Infectious Hematopoietic Necrosis Virus, and Viral Hemorrhagic Septicemia Virus

机译:Hirame弹道病毒,传染性造血坏死病毒和病毒性败血病病毒基因组大小和结构蛋白合成的比较

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摘要

Genomic RNA was extracted from purified virions of hirame rhabdovirus (HRV), infectious hematopoietic necrosis virus (IHNV), and viral hemorrhagic septicemia virus (VHSV). The full-length RNA was analyzed using formaldehyde agarose gel electrophoresis followed by ethidium bromide staining. Compared with an internal RNA size standard, all three viral genomic RNAs appeared to have identical relative mobilities and were estimated to be approximately 10.7 kilobases in length or about 3.7 megadaltons in molecular mass. Structural protein synthesis of HRV, IHNV, and VHSV was studied using cell cultures treated with actinomycin D. At 2h intervals, proteins were labeled with 35S-methionine, extracted, and analyzed by SDS-polyacrylamide gel electrophoresis and autoradiography. The five structural proteins of each of the three viruses appeared in the following order: nucleoprotein (N), matrix protein 1 (M1), matrix protein 2 (M2), glycoprotein (G), and polymerase (L) reflecting both the approximate relative abundance of each protein within infected cells and the gene order within the viral genome.
机译:基因组RNA提取自纯化的hirame弹状病毒(HRV),传染性造血坏死病毒(IHNV)和病毒性败血败血病病毒(VHSV)病毒体。使用甲醛琼脂糖凝胶电泳,然后进行溴乙锭染色,分析全长RNA。与内部RNA大小标准相比,所有三个病毒基因组RNA似乎都具有相同的相对迁移率,估计长度约为10.7千碱基,分子质量约为3.7兆道尔顿。使用放线菌素D处理的细胞培养物研究了HRV,IHNV和VHSV的结构蛋白合成。每隔2小时,用35S-蛋氨酸标记蛋白质,提取并通过SDS-聚丙烯酰胺凝胶电泳和放射自显影进行分析。三种病毒中的每一种的五个结构蛋白按以下顺序出现:核蛋白(N),基质蛋白1(M1),基质蛋白2(M2),糖蛋白(G)和聚合酶(L)反映了两者的相对感染细胞内每种蛋白质的丰度以及病毒基因组内的基因顺序。

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