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狂犬病病毒诱导的BHK21细胞中的RNA合成。

Rabies virus-induced RNA synthesis in BHK21 cells.

作者信息

Holloway B P, Obijeski J F

出版信息

J Gen Virol. 1980 Jul;49(1):181-95. doi: 10.1099/0022-1317-49-1-181.

Abstract

Rabies virus polysomes contained two sizes of messenger RNAs, one of which had a sedimentation value of 30S and another which sedimented at 12 to 16S. RNA extracted from infected cultures contained virion-size RNA, 42S, as well as 30S and 12 to 16S RNA species. Hybridization studies indicated that the 30S and 12 to 16S RNAs had nucleotide sequences which were complementary to virion RNA. RNA. RNA isolated from virus polysomes contained adenylate-rich sequences which were heterogeneous in size and were determined to be about 100 to 250 nucleotides in length on the basis of their migration rates in polyacrylamide gels. Acid-urea agarose gel electrophoresis established that the 30S RNA material was composed of a single RNA species (mol. wt. greater than or equal to 1.65 X 10(6)), whereas the 12 to 16S material could be resolved into at least four distinct species whose mol. wt. ranged from 0.28 to 0.87 X 10(6). When labelled rabies-infected cell RNAs, which were purified by oligo(dT)-cellulose chromatography, were annealed to excess unlabelled virus RNA, digested with ribonuclease T2 and the RNA duplex molecules analysed by polyacrylamide gel electrophoresis, five duplexes could be separated. The mol. wt. of these duplexes were estimated to be 3.2, 1.4, 0.96, 0.55 and 0.39 X 10(6), when compared to the known mol. wt. of vesicular stomatitis virus (VSV) RNA duplexes. This study suggests that the replicative processes of rabies virus are very similar to VSV and that rabies virus proteins are probably translated from smaller than virion-size RNAs.

摘要

狂犬病病毒多核糖体含有两种大小的信使核糖核酸,其中一种沉降值为30S,另一种沉降值为12至16S。从受感染培养物中提取的核糖核酸包含病毒体大小的核糖核酸(42S),以及30S和12至16S的核糖核酸种类。杂交研究表明,30S和12至16S核糖核酸具有与病毒体核糖核酸互补的核苷酸序列。从病毒多核糖体中分离出的核糖核酸含有富含腺苷酸的序列,其大小不均一,根据它们在聚丙烯酰胺凝胶中的迁移率确定其长度约为100至250个核苷酸。酸性尿素琼脂糖凝胶电泳表明,30S核糖核酸物质由单一核糖核酸种类组成(分子量大于或等于1.65×10⁶),而12至16S物质可分解为至少四种不同的种类,其分子量范围为0.28至0.87×10⁶。当通过寡聚(dT)-纤维素柱层析纯化的标记狂犬病感染细胞核糖核酸与过量未标记的病毒核糖核酸退火、用核糖核酸酶T2消化并用聚丙烯酰胺凝胶电泳分析核糖核酸双链分子时,可以分离出五种双链体。与已知分子量的水疱性口炎病毒(VSV)核糖核酸双链体相比,这些双链体的分子量估计为3.2、1.4、0.96、0.55和0.39×10⁶。这项研究表明,狂犬病病毒的复制过程与VSV非常相似,并且狂犬病病毒蛋白可能由小于病毒体大小的核糖核酸翻译而来。

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