Elimination of Mitochondrial DNA fromMammalian CellsNatalya Khozhukhar,1Domenico

Spadafora; Yelitza Rodriguez; Mikhail Alexeyev

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Elimination of Mitochondrial DNA fromMammalian CellsNatalya Khozhukhar,1Domenico

机译：消除Mitococondrial DNA来自Mamalian Cothtnatalya Khozhukhar，1污点

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To cope with DNA damage, mitochondria developed a pathway by whichseverely damaged or unrepairable mitochondrial DNA (mtDNA) moleculesare abandoned and degraded, and new molecules are resynthesized using intacttemplates, if available. In this unit, wedescribe a method that harnesses thispathway to completely eliminate mtDNA from mammalian cells by transientlyoverexpressing the Y147A mutant of human uracil-N-glycosylase (mUNG1).We also provide an alternate protocol for mtDNA depletion using combinedtreatment with ethidium bromide (EtBr) and dideoxycytidine (ddC). Supportprotocols detail approaches for (1) genotyping ρ° cells of human, mouse, andrat origin by PCR; (2) quantitation of mtDNA by quantitative PCR (qPCR);and (3) preparation of calibrator plasmids for mtDNA quantitation.

机译：为了应对DNA损伤，线粒体通过血栓损坏或未分料的线粒体DNA（MTDNA）分子释放和降解而开发了一种途径，并且新分子使用无梗可，如果可用的话，可以使用无梗可。在本机中，呈呈系方法，该方法通过短缺叙述人尿嘧啶-N-糖基酶（Mung1）的Y147A突变体来完全消除来自哺乳动物细胞的MTDNA .We还使用与溴化乙锭（ETBR）的加法处理的MTDNA耗尽提供替代方案和二赤素胞苷（DDC）。 SupportProtocols通过PCR的（1）人，小鼠，Andrat起源的基因分型ρ°细胞的详细方法; （2）定量PCR定量MTDNA（QPCR）;（3）校准质粒的制备用于MTDNA定量。

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《Current Protocols in Cell Biology》 |2018年第1期|共14页
作者
Spadafora; Yelitza Rodriguez; Mikhail Alexeyev;
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作者单位

Department of Physiology and Cell Biology University of South Alabama Mobile Alabama2Flow Cytometry Core Lab University of South Alabama Mobile Alabama;

Department of Physiology and Cell Biology University of South Alabama Mobile Alabama2Flow Cytometry Core Lab University of South Alabama Mobile Alabama;

Department of Physiology and Cell Biology University of South Alabama Mobile Alabama2Flow Cytometry Core Lab University of South Alabama Mobile Alabama;

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原文格式 PDF
正文语种 eng
中图分类细胞生物学;
关键词
cybridsr; mtDNA; mtDNA damager; mtDNA copy numberr; ρ° cell;

机译：糖线;mtdna;mtdna损伤;mtdna拷贝数;ρ°细胞;

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1. Elimination of Mitochondrial DNA fromMammalian CellsNatalya Khozhukhar,1Domenico [J] . Spadafora, Yelitza Rodriguez, Mikhail Alexeyev Current Protocols in Cell Biology . 2018,第1期

机译：消除Mitococondrial DNA来自Mamalian Cothtnatalya Khozhukhar，1污点
2. ELIMINATION OF MUTANT MITOCHONDRIAL GENOMES BY MITOCHONDRIAL-TARGETED DNA EDITING ENZYMES [J] . Moraes Carlos T., Bacman Sandra R., Nissanka Nadee, Molecular genetics and metabolism . 2019,第3期

机译：通过线粒体靶向DNA编辑酶消除突变线粒体基因组
3. ELIMINATION OF MUTANT MITOCHONDRIAL GENOMES BY MITOCHONDRIAL-TARGETED DNA EDITING ENZYMES [J] . Moraes Carlos T., Bacman Sandra R., Nissanka Nadee, Molecular genetics and metabolism . 2019,第3期

机译：通过线粒体靶向DNA编辑酶消除突变线粒体基因组
4. Enhanced Generation of Mitochondrial Reactive Oxygen Species in Cybrids Containing 4977-bp Mitochondrial DNA Deletion [C] . MEI-JIE JOU, TSUNG-I PENG, HONG-YUEH WU, Asian Society for Mitochondrial Research and Medicine . 2005

机译：增强含有4977-BP线粒体DNA缺失的糖线电池活性氧物质的产生
5. Biochemical and genetic characterization of mitochondrial encephalomyopathies resulting from mitochondrial-DNA (mtDNA) and nuclear DNA (nDNA) mutations. [D] . Zheng, Xianxian. 1990

机译：由线粒体DNA（mtDNA）和核DNA（nDNA）突变引起的线粒体脑肌病的生化和遗传特征。
6. The mitochondrial DNA-polymerase promotes elimination of paternal mitochondrial genomes [O] . Zhongsheng Yu, Patrick H. O’Farrell, Nikita Yakubovich, -1

机译：线粒体DNA聚合酶促进消除父系线粒体基因组
7. Maternal inheritance of mitochondrial DNA by diverse mechanisms to eliminate paternal mitochondrial DNA [O] . Sato Miyuki, Sato Ken 2013

机译：母体通过各种机制消除线粒体DNA遗传线粒体DNA

Elimination of Mitochondrial DNA fromMammalian CellsNatalya Khozhukhar,1Domenico

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