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This volume details the most recent advancements in the field of mitochondrial gene expression. Chapters guide readers through methods and protocols on mtDNA replication, transcription, and translation to membrane insertion of the mtDNA-encoded protein products. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, Mitochondrial Gene Expression: Methods and Protocols aims to provide complementary approaches and practical guidelines.
Chapter 7 is available Open Access under a Creative Commons Attribution 4.0 International License via link.springer.com.
1. In vitro Analysis of mtDNA Replication
Jay P. Uhler and Maria Falkenberg
2. In vivo Analysis of mtDNA Replication at the Single Molecule Level and with High Resolution
Marco Tigano, Aaron Fraser Phillips, and Agnel Sfeir
3. In Vitro Reconstitution of Human Mitochondrial Transcription
Azadeh Sarfallah and Dmitry Temiakov
4. Investigating Mitochondrial Transcriptomes and RNA Processing using Circular RNA Sequencing
Irina Kuznetsova, Oliver Rackham, and Aleksandra Filipovska
5. Detection of 5-formylcytosine in Mitochondrial Transcriptome
Lindsey Van Haute and Michal Minczuk
6. Visualization of Mitochondrial RNA Granules in Cultured Cells using 5-Bromouridine Labeling
Vanessa Joanne Xavier and Jean-Claude Martinou
7. Quantitative Proteomics in Drosophila with Holidic Stable Isotope Labeling of Amino Acids in Fruit Flies (SILAF)
Florian A. Schober, Ilian Atanassov, Christoph Freyer, and Anna Wredenberg
8. Mass Spectrometric Analysis of Mitochondrial RNA Modifications
Yuma Ishigami, Tsutomu Suzuki, and Takeo Suzuki
9. mito-?-Seq: A High-throughput Method for Systematic Mapping of Pseudouridine within Mitochondrial RNA
Aldema Sas-Chen, Ronit Nir, and Schraga Schwartz
10. High Throughput Detection of mtDNA Mutations Leading to tRNA Processing Errors
Marita Annika Isokallio and James Bruce Stewart
11. High-throughput Measurement of Mitochondrial RNA Turnover in Human Cultured Cells
Anna V. Kotrys, Lukasz S. Borowski, and Roman J. Szczesny
12. RNA Crosslinking to Analyze the Mitochondrial RNA-binding Proteome
S.L. van Esveld and J.N. Spelbrink
13. Visualizing Mitochondrial Ribosomal RNA and Mitochondrial Protein Synthesis in Human Cell Lines
Matthew Zorkau, Yasmin Proctor-Kent,, Rolando Berlinguer-Palmini, Andrew Hamilton, Zofia M Chrzanowska-Lightowlers, and Robert N. Lightowlers
14. Mitoribosome Profiling From Human Cell Culture - A High Resolution View of Mitochondrial Translation
Pearce Sarah, Cipullo Miriam, Chung Betty, Brierley Ian, and Rorbach Joanna
15. Application of cryo-EM for Visualization of Mitoribosomes
Vivek Singh and A. Amunts
16. Sucrose Gradient Sedimentation Analysis of Mitochondrial Ribosomes
Austin Choi and Antoni Barrientos
17. The Analysis of Yeast Mitochondrial Translation
Andreas Carlström, Magdalena Rzepka, and Martin Ott
18. In situ Studies of Mitochondrial Translation by Cryoelectron Tomography
Robert Englmeier and Friedrich Förster
19. Mitochondrial Complexome Profiling
Heiko Giese, Jana Meisterknecht, Juliana Heidler, and Ilka Wittig
20. Blue Native Electrophoresis to Study the Oxphos Complexes
Erika Fernandez-Vizarra and Massimo Zeviani
21. Identification of Putative Mitochondrial Protease Substrates
Eduard Hofsetz, Pitter F. Huesgen, and Aleksandra Trifunovic
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