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首页> 外文期刊>Journal of Virology >Translation and identification of the viral mRNA species isolated from subcellular fractions of vesicular stomatitis virus-infected cells.
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Translation and identification of the viral mRNA species isolated from subcellular fractions of vesicular stomatitis virus-infected cells.

机译：囊泡口炎病毒感染细胞亚细胞分数中分离的病毒mRNA物种的翻译与鉴定。

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The cytoplasm of vesicular stomatitis virus (VSV)-infected BHK cells has been separated into a fraction containing the membrane-bound polysomes and the remaining supernatant fraction. Total poly(A)-containing RNA was isolated from each fraction and purified. A 17S class of VSV mRNA was found associated almost exclusively with the membrane-bound polysomes, whereas 14,5S and 12S RNAs were found mostly in the postmembrane cytoplasmic supernatant. Poly(A)-containing VSV RNA synthesized in vitro by purified virus was resolved into the same size classes. The individual RNA fractions isolated from VSV-infected cells or synthesized in vitro were translated in cell-free extracts of wheat germ, and their polypeptide products were compared by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. The corresponding in vivo and in vitro RNA fractions qualitatively direct the synthesis of the same viral polypeptides and therefore appear to contain the same mRNA species. By tryptic peptide analysis of their translation products, the in vivo VSV mRNA species have been identified. The 17S RNA, which is compartmentalized on membrane-bound polysomes, codes for a protein of molecular weight 63,000 (P-63) which is most probably a nonglycosylated form of the viral glycoprotein, G. Of the viral RNA species present in the remaining cytoplasmic supernatant, the 14.5S RNA codes almost exclusively for the N protein, whereas the 12S RNA codes predominantly for both the NS and M proteins of the virion.
机译：囊泡口炎病毒（VSV） - 培养的BHK细胞的细胞质已被分离成含有膜结合的多组织的级分和剩余的上清液馏分。从每个馏分中分离出总聚（a）的RNA并纯化。发现17S类的VSV mRNA与膜结合的多肌质几乎完全相关，而最多发现14,5-14,5-14s RNA在后膜细胞质上清液中。通过纯化的病毒在体外合成的VSV RNA占多种VSV RNA，分离成相同的尺寸等级。从VSV感染的细胞中分离的单独RNA部分或在体外合成的小麦胚芽的无细胞提取物，并通过十二烷基硫酸钠 - 聚丙烯酰胺板电泳进行比较它们的多肽产物。体内和体外RNA级分的对应定性地引导相同的病毒多肽的合成，因此似乎含有相同的mRNA物种。通过对其翻译产品的胰蛋白酶肽分析，已经确定了体内VSV mRNA物种。在膜结合的多组织上划分的17℃RNA，用于分子量63,000（p-63）的蛋白质的蛋白质，其最重要的是剩余细胞质中存在的病毒糖蛋白的核化形式的病毒糖蛋白。上清液，几乎专门针对N蛋白的14.5s RNA码，而12S RNA码主要针对病毒夫岛的NS和M蛋白。

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