Mori T, Wakabayashi Y, Takagi S
J Biochem. 1978 Nov;84(5):1103-11. doi: 10.1093/oxfordjournals.jbchem.a132224.
Poly(A)-containing RNA has been isolated from the cotyledons of soybean seeds by adsorption on a poly(U)-Sepharose column. Approximately 0.15% of the total soybean RNA applied bound to the column. The bound RNA (poly(A)-containing RNA) was shown to be mRNA by its ability to serve as template in a cell-free system derived from wheat germ. Poly(A)-containing RNA was polydisperse, migrating from approximately 50,000 to 700,000 daltons with a mean of 150,000 daltons in polyacrylamide gel electrophoresis. The size of the poly(A) portion of this RNA was in the range of 55 to 290 nucleotides. The adenylic acid content of the presumed poly(A) fragment was about 95%. The radioactive products of translation directed by the poly(A)-containing RNA in the wheat germ cell-free system were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and by immunoprecipitation using antisera against beta-conglycinin and glycinin. The results of this investigation show that mRNAs for the subunit proteins of the major components of a soybean storage protein exist in the poly(A)-containing RNA preparation obtained from the cotyledons of dry soybean seeds.
通过吸附在聚(U)-琼脂糖柱上,已从大豆种子的子叶中分离出含聚(A)的RNA。所加样的大豆总RNA中约0.15%与柱结合。结合的RNA(含聚(A)的RNA)通过其在源自小麦胚芽的无细胞系统中作为模板的能力被证明是mRNA。含聚(A)的RNA是多分散的,在聚丙烯酰胺凝胶电泳中迁移率约为50,000至700,000道尔顿,平均为150,000道尔顿。该RNA的聚(A)部分大小在55至290个核苷酸范围内。推测的聚(A)片段的腺苷酸含量约为95%。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及使用针对β-伴大豆球蛋白和大豆球蛋白的抗血清进行免疫沉淀,分析了含聚(A)的RNA在小麦胚芽无细胞系统中指导的翻译放射性产物。这项研究的结果表明,从干大豆种子子叶获得的含聚(A)的RNA制剂中存在大豆贮藏蛋白主要成分的亚基蛋白的mRNA。