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水疱性口炎病毒信使核糖核酸种类的核苷酸序列复杂性、分子量及聚腺苷酸含量

Nucleotide sequence complexities, molecular weights, and poly(A) content of the vesicular stomatitis virus mRNA species.

作者信息

Rose J K, Knipe D

出版信息

J Virol. 1975 Apr;15(4):994-1003. doi: 10.1128/JVI.15.4.994-1003.1975.

Abstract

Poly(A)-containing vesicular stomatitis virus mRNA species synthesized in vesicular stomatitis virus-infected cells have been separated into four bands by electrophoresis on formamide-polyacrylamide gels. Two-dimensional fingerprints of ribonuclease T-1 and ribonuclease A digests of the RNA from each band show that they contain unique oligonucleotide sequences as well as 60 to 125 nucleotides of poly(A). The fingerprints were used to determine the nucleotide sequence complexities of RNA from three of the bands. Two contain nucleotide sequences which account completely for their molecular weights (0.70 times 10-6 and 0.55 times 10-6) determined by gel electrophoresis and sedimentation rate, and, therefore, these are radiochemically pure RNA species. The most rapidly migrating band must contain two ro three different RNA species since it has a molecular weight of 0.28 times 10-6, determined by physical methods, and a nucleotide sequence complexity two to three times that expected for a pure RNA species of this size. These data are in complete accord with translational studies (accompanying paper) which show that each of the two pure RNA species codes for a distinct viral protein, whereas the third codes for two viral proteins. From the molecular weight and sequence complexity determinations on mRNA from the bands, we conclude that most of the vesicular stomatitis virus genome is transcribed into discrete mRNA species.

摘要

在水疱性口炎病毒感染的细胞中合成的含聚腺苷酸的水疱性口炎病毒信使核糖核酸种类,通过在甲酰胺-聚丙烯酰胺凝胶上电泳已被分离成四条带。对每条带的核糖核酸经核糖核酸酶T-1和核糖核酸酶A消化后的二维指纹图谱显示,它们含有独特的寡核苷酸序列以及60至125个聚腺苷酸核苷酸。这些指纹图谱被用于确定其中三条带的核糖核酸的核苷酸序列复杂性。其中两条带含有完全能解释其通过凝胶电泳和沉降速率测定的分子量(分别为0.70×10⁻⁶和0.55×10⁻⁶)的核苷酸序列,因此,这些是放射化学纯的核糖核酸种类。迁移最快的带必定包含两种或三种不同的核糖核酸种类,因为通过物理方法测定其分子量为0.28×10⁻⁶,而其核苷酸序列复杂性是这种大小的纯核糖核酸种类预期的两到三倍。这些数据与翻译研究(随附论文)完全一致,该研究表明两种纯核糖核酸种类各自编码一种独特的病毒蛋白,而第三种编码两种病毒蛋白。从对这些带的信使核糖核酸的分子量和序列复杂性测定结果,我们得出结论,水疱性口炎病毒基因组的大部分被转录成离散的信使核糖核酸种类。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/882c/354544/b90539470672/jvirol00232-0333-a.jpg

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