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黑麦胚系统作为小麦系统的替代物用于体外蛋白质合成。

The rye embryo system as an alternative to the wheat system for protein synthesis in vitro.

作者信息

Carlier A R, Peumans W J

出版信息

Biochim Biophys Acta. 1976 Nov 1;447(4):436-44. doi: 10.1016/0005-2787(76)90081-2.

Abstract

Isolated rye embryos are a readily available source for the preparation of very active, cell-free, protein-synthesizing systems. Incorporation levels up to 2000 pmol leucine per 50 mul assay are routinely obtained at saturating TMV (Tobacco mosaic virus) RNA concentrations; at limiting messenger RNA concentrations the incorporation exceeds 1000 leucine molecules per TMV RNA molecule. The characteristics of this cell-free system for the translation of TMV RNA are identical with those of a similarly prepared wheat germ system. The major advantage of the rye embryo system is its high reliability as compared to the unpredictable wheat germ system. Sucrose gradient analysis of the reaction mixture during the incubation shows an extensive polysome formation with TMV RNA and demonstrates efficient polypeptide chain release.

摘要

分离的黑麦胚是制备活性极高的无细胞蛋白质合成系统的现成来源。在饱和烟草花叶病毒(TMV)RNA浓度下,每50微升测定中亮氨酸掺入水平通常可达2000皮摩尔;在信使RNA浓度受限的情况下,每一个TMV RNA分子的亮氨酸掺入量超过1000个亮氨酸分子。这个用于翻译TMV RNA的无细胞系统的特性与用类似方法制备的小麦胚系统相同。与难以预测的小麦胚系统相比,黑麦胚系统的主要优点是其高可靠性。孵育过程中对反应混合物进行蔗糖梯度分析显示,TMV RNA能形成大量多核糖体,并证明有高效的多肽链释放。

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