INJECTION OF MITOCHONDRIA INTO HUMAN-CELLS LEADS TO A RAPID REPLACEMENT OF THE ENDOGENOUS MITOCHONDRIAL-DNA
INJECTION OF MITOCHONDRIA INTO HUMAN-CELLS LEADS TO A RAPID REPLACEMENT OF THE ENDOGENOUS MITOCHONDRIAL-DNA
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DOI：
10.1016/0092-8674(88)90423-0
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发表时间：
1988-03-25
期刊：
影响因子：
64.5
通讯作者：
ATTARDI, G
中科院分区：
文献类型：
--
作者：
KING, MP;ATTARDI, G
将含有编码氯霉素抗性标记的线粒体DNA（mtDNA）的分离的人线粒体注射到来自两种不同的人敏感细胞系143 BTK-和HT 1080 - 6 TG的细胞中。根据有关哺乳动物细胞中引入体积耐受性的现有证据，估计平均而言，每个细胞中引入不到一个微粒子。在选择性条件下，线粒体在受体细胞中以大于2-3倍的频率建立。10-3.多个mtDNA和核DNA多态性的分析表明，外来mtDNA的快速替换的居民mtDNA。显微注射后6至10周，除了一个HT 1080 - 6 TG转化体之外，所有的HT 1080 - 6 TG转化体都完成了这种替换，并且在大多数143个BTK-转化体中几乎完成。在选择的非常早期阶段的转化体的mtDNA的定量行为强烈地表明，细胞内的mtDNA选择在这种替换中发挥了至关重要的作用，与线粒体遗传学的重大影响。
Isolated human mitochondria containing a mitochondrial DNA (mtDNA) coded chloramphenicol resistance marker were injected into cells from two different human sensitive cell lines, 143BTK- and HT1080-6TG, which had been partially depleted of their mtDNA by ethidium bromide treatment. On the basis of the available evidence concerning the tolerance of introduced volumes into mammalian cells, it is estimated that, on the average, less than one mitochondrion was introduced into each cell. Under selective conditions, the mitochondria became established in the recipient cells with a frequency greater than 2-3 .times. 10-3. An analysis of multiple mtDNA and nuclear DNA polymorphisms revealed a rapid replacement of the resident mtDNA by the exogenous mtDNA. Six to ten weeks after microinjection, this replacement was complete in all but one of the HT1080-6TG transformants, and nearly complete in the majority of the 143BTK- transformants. The quantitative behavior of the mtDNA of the transformants at very early stages of selection strongly suggests that intracellular mtDNA selection played a crucial role in this replacement, with significant implications for mitochondrial genetics.