Isolation and characterization of mitochondrial nucleoids from the yeast Pichia jadinii.
Isolation and characterization of mitochondrial nucleoids from the yeast Pichia jadinii.
复制标题
酵母 Pichia jadinii 中线粒体核的分离和表征。
DOI：
10.1093/oxfordjournals.pcp.a029017
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发表时间：
1996
影响因子：
4.9
通讯作者：
N. Sando
中科院分区：
文献类型：
--
作者：
I. Miyakawa;C. Okazaki;T. Higashi;Y. Furutani;N. Sando
从酵母 Pichia jadinii 中以高纯度分离出线粒体 (mt) 核，其中线粒体 DNA (mtDNA) 是线性的。场反转凝胶电泳 (FIGE) 显示，可以从分离的 mt 核仁中完整分离出大量 mtDNA，为 41 kbp 的线性分子。在 mt 核部分中检测到 15 种不同的蛋白质，其中 8 种蛋白质与 DNA 结合。在 SDS 存在下进行凝胶电泳后，mt 核蛋白和 DNA 结合蛋白的模式与来自酿酒酵母的此类蛋白质的模式有些不同。从毕赤酵母属和威利奥普属酵母的其他四种酵母中分离出的相应蛋白质在 SDS 存在下的电泳迁移率也彼此不同。在免疫印迹实验中，针对酿酒酵母 mt 核仁中 67 kDa 蛋白产生的抗体和对酿酒酵母 mt 核仁中 48 kDa 蛋白具有特异性的 YMN-1 单克隆抗体，无法识别来自 Pichia jadinii 和其他四种酵母的 mt 核仁中的任何蛋白质。结果表明酵母 mt 核中的蛋白质具有相当大的多样性。
Mitochondrial (mt) nucleoids were isolated with a high degree of purity from the yeast Pichia jadinii, in which the mitochondrial DNA (mtDNA) is linear. Field-inversion gel electrophoresis (FIGE) revealed that significant amounts of mtDNA could be isolated intact, as linear molecules of 41 kbp, from the isolated mt-nucleoids. Fifteen different proteins were detected in the mt-nucleoid fraction and, eight of these proteins bound to DNA. The patterns of mt-nucleoid proteins and of the DNA-binding proteins after gel electrophoresis in the presence of SDS were somewhat different from those of such proteins from Saccharomyces cerevisiae. The corresponding proteins isolated from the mt-nucleoids of four other species of yeast in the genera Pichia and Williopsis also differed from one another in terms of electrophoretic mobility in the presence of SDS. In immunoblotting experiments, antibodies that had been raised against the 67-kDa protein of mt-nucleoids from S.cerevisiae and the YMN-1 monoclonal antibody that is specific for a 48-kDa protein in the mt-nucleoids from S. cerevisiae did not recognize any proteins in the mt-nucleoids from Pichia jadinii and four other species of yeast. The results suggest the considerable diversity of the proteins in the mt-nucleoids of yeasts.