Dai H, Lo Y S, Wu C Y, Tsou C L, Hsu G S, Chern C G, Ruddat M, Chiang K S
Institute of Botany, Academia Sinica, Taipei, Taiwan, Republic of China.
Plant Physiol. 1991 May;96(1):319-23. doi: 10.1104/pp.96.1.319.
For studies of in organello mitochondrial protein synthesis in rice, Oryza sativa L., conventional surface-sterilization procedures were demonstrated to be ineffective. Because of the over-whelmingly efficient [(35)S]methionine utilization by contaminating bacteria, even "essentially bacteria-free" rice mitochondria were shown to be unsuitable for the study of in organello protein synthesis. We developed a procedure to obtain a bacteria-free preparation of rice mitochondria. Such mitochondria favored a membrane-dependent ATP-generating system over an external ATP-generating system as the energy supplement for in organello protein synthesis. Two distinct classes of [(35)S]methionine-labeled, cycloheximide-insensitive products were detected: an electrophoretically unresolved population and a set of some 22 to 27 discrete polypeptide species, each with a characteristic electrophoretic mobility and relative abundance.
对于水稻（Oryza sativa L.）线粒体在细胞器内蛋白质合成的研究，传统的表面灭菌程序被证明是无效的。由于污染细菌对[³⁵S]甲硫氨酸的利用效率极高，即使是“基本无细菌”的水稻线粒体也被证明不适用于细胞器内蛋白质合成的研究。我们开发了一种获得无细菌水稻线粒体制剂的程序。这种线粒体更倾向于将依赖膜的ATP生成系统而非外部ATP生成系统作为细胞器内蛋白质合成的能量补充。检测到两类不同的[³⁵S]甲硫氨酸标记的、对环己酰亚胺不敏感的产物：一类是电泳未分离的群体，另一类是一组约22至27种离散的多肽物种，每种都具有独特的电泳迁移率和相对丰度。
