Laboratorium voor Plantenbiochemie, K.U. Leuven, Vaartstraat 24, B-3000, Leuven, Belgium.
Planta. 1977 Jan;136(3):195-201. doi: 10.1007/BF00385985.
Extracts prepared from dry wheat (Triticum aestivum L.; var. Cama) and rye (Secale cereale L.; var. Celestijner) embryos exhibit relatively high endogenous messenger activities in an in vitro protein synthesizing system: up to 400 pmol leucine are incorporated per 50 μl reaction mixture; the messenger/ribosome ratio is estimated at about 0.03. Sucrose gradient analysis of the incubation mixture during cellfree protein synthesis shows a progressive in vitro polysome formation with the native endogenous mRNPs. The translation of these messengers, although dependent on initiation, follows linear kinetics. The native mRNP particles can be separated on sucrose gradients in 6 distinct size classes with sedimentation values varying from 25 S to 104 S. Their size is strongly reduced after deproteinization so that the naked mRNAs sediment in a range between 8 S and 35 S. The molecular weights of the proteins synthesized under direction of the native RNA particles range from 12,000 to 70,000 as estimated by polyacrylamide gel electrophoresis.
从干小麦(Triticum aestivum L.;var. Cama)和黑麦(Secale cereale L.;var. Celestijner)胚中提取的物质在体外蛋白质合成系统中表现出相对较高的内源性信使活性:每个 50μl 反应混合物中掺入多达 400pmol 亮氨酸;信使/核糖体的比例估计约为 0.03。在无细胞蛋白质合成过程中对孵育混合物的蔗糖梯度分析显示出与天然内源性 mRNP 一致的体外多核糖体形成的进行性过程。尽管这些信使的翻译依赖于起始,但遵循线性动力学。天然 mRNP 颗粒可以在蔗糖梯度上分离成 6 个不同的大小类,沉降值从 25S 到 104S 不等。脱蛋白后,它们的大小大大减小,使得裸 mRNA 在 8S 到 35S 之间的范围内沉降。根据聚丙酰胺凝胶电泳估计,在天然 RNA 颗粒指导下合成的蛋白质的分子量范围从 12,000 到 70,000。