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Expression of bacterial endonucleases in Saccharomyces cerevisiae mitochondria
Mitochondrion ( IF 4.4 ) Pub Date : 2002-11-01 , DOI: 10.1016/s1567-7249(02)00037-5
Sarah L Donahue 1 , Uma Lakshmipathy , Colin Campbell
Affiliation  

Expression vectors were created in which the 5' end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.

中文翻译:

细菌核酸内切酶在酿酒酵母线粒体中的表达

创建表达载体,其中编码线粒体靶向肽的酿酒酵母 CDC9 基因的 5' 端与 EcoR I、Hind III 和 Pst I 核酸内切酶基因的编码区框内克隆。EcoR I 和 Hind III 融合蛋白的表达抑制了酵母在含甘油培养基上的生长,并导致其线粒体 DNA 的几乎定量限制性消化。相比之下,不识别酵母线粒体 DNA 内任何位点的 Pst I 的表达对含甘油培养基中的生长没有影响,也不影响线粒体基因组的完整性。
更新日期:2002-11-01
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