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首页> 外文期刊>Journal of Virology >Cell-free translation of RNA synthesized in vitro by a transcribing nucleoprotein complex prepared from purified vesicular stomatitis virus.
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Cell-free translation of RNA synthesized in vitro by a transcribing nucleoprotein complex prepared from purified vesicular stomatitis virus.

机译：通过纯化的脉湿性口炎病毒制备的转录核蛋白复合物在体外合成的RNA的无细胞翻译。

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摘要

The RNA species synthesized in vitro by a transcribing nucleoprotein (TNP) complex of vesicular stomatitis virus (VSV) were translated with high efficiency in a fractionated cell-free system derived from reticulocytes. The use of TNP complexes isolated from VSV Indiana, VSV New Jersey, and Chandipura viruses showed that in each case the predominant polypeptides synthesized had electrophoretic mobilities identical to their virion N, NS, and M polypeptides in proportions reflecting those found in infected cells rather than purified virions. A minor polypeptide corresponding to unglycosylated polypeptide G was also observed, but the in vitro synthesis of polypeptide L was not detected. The addition of RNase inhibitor to transcription mixtures markedly increased the rate of RNA synthesis. Furthermore, the messenger activity of the RNA was significantly enhanced. The inclusion of S-adenosyl L-methionine during transcription substantially increased the messenger activity of the product RNA, suggesting a requirement for methylation. Fractionation by oligodeoxythymidylic acid-cellulose chromatography revealed that the RNA required a polyadnylic acid tract for messenger activity.
机译：通过转录核开蛋白（TNP）在体外合成的RNA物种在衍生自网状细胞的分级无细胞系统中以高效率转化为脉络膜炎病毒（VSV）。从VSV印第安纳州分离的TNP复合物和Chandipura病毒的使用表明，在每种情况下，合成的主要多肽具有与其病毒尼，NS和M多肽相同的电泳迁移率，其比例反映在感染细胞中发现的那些纯化的病毒粒子。还观察到对应于非糖基化多肽G的次要多肽，但未检测到多肽L的体外合成。添加RNase抑制剂对转录混合物显着增加了RNA合成率。此外，RNA的信使活动显着提高。在转录过程中包含S-腺苷L-蛋氨酸显着增加了产物RNA的信使活性，表明甲基化的要求。通过寡脱氧脂亚酸纤维素色谱分离的分级显示，RNA需要用于信使活动的多酰基酸流。

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