Mitochondrial DNA : methods and protocols / edited by Jeffrey A. Stuart. — Dordrecht ; London : Humana Press, c2009. – (58.17/M592/v.554) |
Contents
Contents
Preface
Contributors
Part I: MITOCHONDRIAL DNA REPLICATION, TRANSCRIPTION, TRANSLATION, AND RESPIRATORY COMPLEX ASSEMBLY
1. Biochemical Isolation of mtDNA Nucleoids from Animal Cells.
2. Analysis of Mitochondrial DNA by Two-Dimensional Agarose Gel Electrophoresis
3. Comparative Purification Strategies for Drosophila and Human Mitochondrial DNA Replication Proteins: DNA Polymerase γ and Mitochondrial Single-Stranded DNA-Binding Protein
4. Functional Analysis of Mutant Mitochondrial DNA Polymerase Proteins Involved in Human Disease
5. Preparation of Human Mitochondrial Single-Stranded DNA-Binding Protein
6. Methods for Assessing Binding of Mitochondrial Transcription Factor A (TFAM) to DNA
7. Inducible Expression in Human Cells, Purification, and In Vitro Assays for the Mitochondrial DNA Helicase Twinkle
8. Purification Strategy for Recombinant Forms of the Human Mitochondrial DNA Helicase
9. Methods for Studying Mitochondrial Transcription Termination with Isolated Components
10. Oxidative Phosphorylation: Synthesis of Mitochondrially Encoded Proteins and Assembly of Individual Structural Subunits into Functional Holoenzyme Complexes
Part II: MITOCHONDRIAL DNA DAMAGE AND REPAIR11. Reactive Oxygen Species Production by Mitochondria.
12. Measuring mtDNA Damage Using a Supercoiling-Sensitive qPCR Approach
13. Quantitative Analysis of Oxidized Guanine, 8-Oxoguanine, in Mitochondrial DNA by Immunofluorescence Method 199
14. In Vitro Measurement of DNA Base Excision Repair in Isolated Mitochondria 213
15. Targeting Repair Proteins to the Mitochondria of Mammalian Cells Through Stable Transfection, Transient Transfection, Viral Transduction, and TAT-Mediated Protein Transduction 233
16. Construction and Characterization of a Cell Line Deficient in Repair of Mitochondrial, but Not Nuclear, Oxidative DNA Damage 251
Part III: MITOCHONDRIAL DNA MUTATIONS
17. Mitochondrial DNA Oxidative Damage and Mutagenesis in Saccharomyces cerevisiae 267
18. Determination of DNA Mutation Load in Human Tissues Using Denaturing HPLC-Based Heteroduplex Analysis 287
19. Rapid Identification of Unknown Heteroplasmic Mitochondrial DNA Mutations with Mismatch-Specific Surveyor Nuclease 301
20. Collection of Isolated Cells for Studying Mitochondrial DNA Mutations Within Individual Cells 315
21. Quantitative Analysis of Somatic Mitochondrial DNA Mutations by Single-Cell Single-Molecule PCR 329
22. Measuring DNA Precursor Pools in Mitochondria
23. Establishment of Human Cell Lines Lacking Mitochondrial DNA