人线粒体DNA含量的Southern印迹和非放射性探针杂交分析

Joel H. Wheeler, Carolyn K. J. Young, Matthew J. Young
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引用次数: 9

摘要

单个细胞可以包含数千个线粒体DNA基因组或mtDNA的拷贝。评估mtDNA含量的工具对于临床和毒理学研究是必要的,因为mtDNA耗竭与遗传疾病和药物毒性有关。例如,mtDNA耗竭综合征是典型的致命性儿童疾病,其特征是受影响组织中mtDNA含量严重下降。在某些核苷逆转录酶抑制剂治疗的人类免疫缺陷病毒感染患者中也有线粒体毒性和mtDNA耗损的报道。此外,细胞培养研究表明,暴露于氧化应激刺激mtDNA降解。在这里,我们概述了一种Southern blot和非放射性地高辛标记探针杂交方法来估计人类基因组DNA样本中的mtDNA含量。©2019 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Analysis of Human Mitochondrial DNA Content by Southern Blotting and Nonradioactive Probe Hybridization

A single cell can contain several thousand copies of the mitochondrial DNA genome or mtDNA. Tools for assessing mtDNA content are necessary for clinical and toxicological research, as mtDNA depletion is linked to genetic disease and drug toxicity. For instance, mtDNA depletion syndromes are typically fatal childhood disorders that are characterized by severe declines in mtDNA content in affected tissues. Mitochondrial toxicity and mtDNA depletion have also been reported in human immunodeficiency virus–infected patients treated with certain nucleoside reverse transcriptase inhibitors. Further, cell culture studies have demonstrated that exposure to oxidative stress stimulates mtDNA degradation. Here we outline a Southern blot and nonradioactive digoxigenin-labeled probe hybridization method to estimate mtDNA content in human genomic DNA samples. © 2019 by John Wiley & Sons, Inc.

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