Institut de Biochimie Cellulaire et Neurochimie du CNRS, 1 rue Camille Saint-Saëns, 33077, Bordeaux Cedex, France.
Plant Mol Biol. 1983 Jul;2(4):167-75. doi: 10.1007/BF01578376.
DNA synthesis was studied using purified wheat embryo mitochondria as well as mitochondrial lysates deprived of endogenous DNA. The optimal conditions for DNA synthesis are very similar in both systems: ATP stimulates dramatically mitochondrial DNA synthesis and magnesium is a better co-factor than manganese, contrary to what has been reported in animal mitochondrial systems. Wheat mitochondrial DNA synthesis is resistant to aphidicolin and strongly inhibited by dideoxythymidine triphosphate and ethidium bromide. Thus, the DNA polymerase involved in this system seems to be the same as that previously purified and characterized from wheat embryo mitochondria (Christopheet al., Plant Science Letters 21: 181, 1981). Two different approaches: restriction endonuclease digestion followed by electrophoresis, and autoradiography and cesium chloride equilibrium centrifugation of mitochondrial DNA, where BrdUTP has been incorporated instead of TTP, show that long stretches of the mitochondrial genome have been synthesized.
使用纯化的小麦胚线粒体以及缺乏内源性 DNA 的线粒体裂解物研究了 DNA 合成。在这两种系统中,DNA 合成的最佳条件非常相似:ATP 显著刺激线粒体 DNA 合成,并且镁是比锰更好的辅助因子,与动物线粒体系统中的报道相反。小麦线粒体 DNA 合成对 aphidicolin 具有抗性,并被双脱氧胸苷三磷酸和溴化乙锭强烈抑制。因此,参与该系统的 DNA 聚合酶似乎与先前从小麦胚线粒体中纯化和表征的酶相同(Christopheet al.,Plant Science Letters 21: 181, 1981)。两种不同的方法:用限制酶消化,然后进行电泳,以及用 BrdUTP 代替 TTP 掺入的线粒体 DNA 的放射自显影和铯氯化物平衡离心,显示已合成了线粒体基因组的长片段。