Mitochondrial DNA : methods and protocols / edited by Jeffrey A. Stuart.

Format:

Book

Contributor:

Stuart, Jeffrey A. (Jeffrey Alan), 1968-

Series:

Methods in molecular biology (Clifton, N.J.) 1064-3745 ; v. 554.

Methods in molecular biology, 1064-3745 ; v. 554

Language:

English

Subjects (All):

Mitochondrial DNA--Laboratory manuals.

Mitochondrial DNA.

DNA, Mitochondrial.

Genetic Techniques.

Medical Subjects:

DNA, Mitochondrial.

Genetic Techniques.

Genre:

Laboratory manuals.

Physical Description:

xi, 403 pages : illustrations (1 color) ; 27 cm.

Edition:

Second edition.

Place of Publication:

New York : Humana Press, [2009]

Summary:

Since the publication of the first edition, the number of unique heritable mtDNA mutations recognized as being associated with bioenergetic dysfunction, cell death and disease has grown. Likewise, our understanding of the basic biology of somatic mtDNA mutations continues to improve. In Mitochondrial DNA: Methods and Protocols, Second Edition, specialists from eight countries share their expertise, providing detailed protocols for studying many aspects of mtDNA. The volume is divided into three sections, which cover the transduction of information from mtDNA to functionally active respiratory complexes, mitochondrial reactive oxygen species (ROS) production as well as mtDNA damage and its repair, and the identification and quantification of heteroplasmic mtDNA mutations. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.

Comprehensive and authoritative, Mitrochondrial DNA: Methods and Protocols, Second Edition promises to aid researchers in further expanding our knowledge of this vital area of cell biology.

Contents:

Part I Mitochondrial DNA Replication, Transcription, Translation, and Respiratory Complex Assembly

1 Biochemical Isolation of mtDNA Nucleoids from Animal Cells / Daniel F. Bogenhagen 3

2 Analysis of Mitochondrial DNA by Two-Dimensional Agarose Gel Electrophorsis / Aurelio Reyes, Takehiro Yasukawa, Tricia J. Cluett, Ian J. Holt 15

3 Comparative Purification Strategies for Drosophila and Human Mitochondrial DNA Replication Proteins: DNA Polymerase γ and Mitochondrial Single-Stranded DNA-Binding Protein / Marcos T. Oliveira, Laurie S. Kaguni 37

4 Functional Analysis of Mutant Mitochondrial DNA Polymerase Proteins Involved in Human Disease / Sherine S. L. Chan, William C. Copeland 59

5 Preparation of Human Mitochondrial Single-Stranded DNA-Binding Protein / Matthew J. Longley, Leslie A. Smith, William C. Copeland 73

6 Methods for Assessing Binding of Mitochondrial Transcription Factor A (TFAM) to DNA / Atsushi Fukuoh, Dongchon Kang 87

7 Inducible Expression in Human Cells, Purification, and In Vitro Assays for the Mitochondrial DNA Helicase Twinkle / Steffi Goffart, Hans Spelbrink 103

8 Purification Strategy for Recombinant Forms of the Human Mitochondrial DNA Helicase / Tawn D. Ziebarth, Laurie S. Kaguni 121

9 Methods for Studying Mitochondrial Transcription Termination with Isolated Components / Paola Loguercio Polosa, Stefania Deceglie, Marina Roberti, Maria Nicola Gadaleta, Palmiro Cantatore 127

10 Oxidative Phosphorylation: Synthesis of Mitochondrially Encoded Proteins and Assembly of Individual Structural Subunits into Functional Holoenzyme Complexes / Scot C. Leary, Florin Sasarman 143

Part II Mitochondrial DNA Damage And Repair

11 Reactive Oxygen Species Production by Mitochondria / Andrian J. Lambert, Martin D. Brand 165

12 Measuring mtDNA Damage Using a Supercoiling-Sensitive qPCR Approach / Sam W. Chan, Junjian Z. Chen 183

13 Quantitative Analysis of Oxidized Guanine, 8-Oxoguanine, in Mitochondrial DNA by Immunofluorescence Method / Mizuki Ohno, Sugako Oka, Yusaku Nakabeppu 199

14 In Vitro Measurement of DNA Base Excision Repair in Isolated Mitochondria / Melissa M. Page, Jeffrey A. Stuart 213

15 Targeting Repair Proteins to the Mitochondria of Mammalian Cells Through Stable Transfection, Transient Transfection, Viral Transduction, and TAT-Mediated Protein Transduction / Christopher A. Koczor, Janet W. Snyder, Inna N. Shokolenko, Allison W. Dobson, Glenn L. Wilson, Susan P. LeDoux 233

16 Construction and Characterization of a Cell Line Deficient in Repair of Mitochondrial, but Not Nuclear, Oxidative DNA Damage / Sugako Oka, Mizuki Ohno, Yusaku Nakabeppu 251

Part III Mitochondrial DNA Mutations

17 Mitochondrial DNA Oxidative Damage and Mutagenesis in Saccharomyces cerevisiae / Lyra M. Griffiths, Nicole A. Doudican, Gerald S. Shadel, Paul W. Doetsch 267

18 Determination of DNA Mutation Load in Human Tissues Using Denaturing HPLC-Based Heteroduplex Analysis / Kok Seong Lim, Robert K. Naviaux, Richard H. Haas 287

19 Rapid Identification of Unknown Heteroplasmic Mitochondrial DNA Mutations with Mismatch-Specific Surveyor Nuclease / Sylvie Bannwarth, Vincent Procaccio, Véronique Paquis-Flucklinger 301

20 Collection of Isolated Cells for Studying Mitochondrial DNA Mutations Within Individual Cells / Yevgenya Kraytsberg, Natalya Bodyak, Susan Myerow, Alexander Nicholas, Konstantin Ebralidze, Konstantin Khrapko 315

21 Quantitative Analysis of Somatic Mitochondrial DNA Mutations by Single-Cell Single-Molecule PCR / Yevgenya Kraytsberg, Natalya Bodyak, Susan Myerow, Alexander Nicholas, Konstantin Ebralidze, Konstantina Khrapko 329

22 Measuring DNA Precursor Pools in Mitochondria / Christopher K. Matthews, Linda J. Wheeler 371

23 Establishment of Human Cell Lines Lacking Mitochondrial DNA / Kazunari Hashiguchi, Qiu-Mei Zhang-Akiyama 383.

Notes:

"Springer protocols"--Cover.

Includes bibliographical references and index.

ISBN:

9781934115602

1934115606

9781597455213

1597455210

OCLC:

436968412