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小麦胚核糖核酸。十四。从小麦胚芽中大规模分离信使核糖核酸及其翻译能力与吸胀小麦胚信使核糖核酸的比较。

Wheat embryo ribonucleates. XIV. Mass isolation of mRNA from wheat germ and comparison of its translational capacity with that of mRNA from imbibing wheat embryos.

作者信息

Cuming A C, Kennedy T D, Lane B G

出版信息

Can J Biochem. 1979 Sep;57(9):1170-5. doi: 10.1139/o79-151.

Abstract

Commercially milled wheat germ is shown to be a convenient source material for facile recovery of mass (milligram) quantities of highly purified poly(A)-rich RNA. This poly(A)-rich RNA is efficiently translated in a nuclease-treated extract of rabbit reticulocytes. By sucrose density gradient fractionation of bulk poly(A)-rich RNA from wheat germ, it has been possible to show that there is a direct relationship between the molecular weights of the polypeptide products of cell-free synthesis and the molecular weights of the wheat mRNA molecules which program their synthesis. As assessed by SDS -- polyacrylamide gel electrophoresis, the same array of polypeptides is synthesized when nuclease-treated reticulocyte extract is programmed by poly(A)-rich RNA from either commercially supplied or laboratory-prepared wheat embryos. Significantly, there are gross quantitative if not qualitative differences between the translational capacities of poly(A)-rich RNA from dry and imbibing wheat embryos, and the possible importance of these differences for interpreting a changing pattern of polypeptide synthesis in imbibing wheat embryos is the subject of a brief discussion.

摘要

商业研磨的小麦胚芽被证明是一种方便的原料,可用于轻松回收毫克量的高度纯化的富含多聚腺苷酸(poly(A))的RNA。这种富含poly(A)的RNA在经核酸酶处理的兔网织红细胞提取物中能高效翻译。通过对小麦胚芽中大量富含poly(A)的RNA进行蔗糖密度梯度分级分离,已能够表明无细胞合成的多肽产物的分子量与编码其合成的小麦mRNA分子的分子量之间存在直接关系。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-聚丙烯酰胺凝胶电泳)评估,当用来自商业供应或实验室制备的小麦胚的富含poly(A)的RNA对经核酸酶处理的网织红细胞提取物进行编程时,会合成相同系列的多肽。值得注意的是,干燥和吸胀小麦胚的富含poly(A)的RNA的翻译能力之间存在显著的数量差异(如果不是质量差异的话),并且这些差异对于解释吸胀小麦胚中多肽合成模式变化的可能重要性是简短讨论的主题。

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