Aarhus University Seal

Measuring the Activity of DNA Repair Enzymes in Isolated Mitochondria

Research output: Contribution to book/anthology/report/proceedingBook chapterResearchpeer-review

Nuclear, mitochondrial and plastidic DNA is constantly exposed to conditions, such as ultraviolet radiation or reactive oxygen species, which will induce chemical modifications to the nucleotides. Unless repaired, these modifications can lead to mutations, so the nucleus, mitochondria and plastids each contains a number of DNA repair systems. We here describe assays for measuring the enzyme activities associated with the base-excision repair pathway in potato tuber mitochondria. As the name implies, this pathway involves removing a modified base and replacing it with an undamaged base. Activity of each of the enzymes involved, DNA glycosylase, apurinic/apyrimidinic endonuclease, DNA polymerase and DNA ligase can be measured by incubating a mitochondrial extract with a specifically designed oligonucleotide. After incubation, the reaction mixture is separated on a polyacrylamide gel, and the amounts of specific products formed is estimated by autoradiography, which makes it possible to calculate the enzymatic activity.

Original languageEnglish
Title of host publicationPlant Mitochondria
Place of publicationNew York
PublisherHumana Press
Publication year2022
ISBN (print)978-1-0716-1652-9
ISBN (electronic)978-1-0716-1653-6
Publication statusPublished - 2022
SeriesMethods in Molecular Biology

Bibliographical note

Publisher Copyright:
© 2022, Springer Science+Business Media, LLC, part of Springer Nature.

    Research areas

  • Base excision repair, DNA repair, mtDNA, Plant mitochondria

See relations at Aarhus University Citationformats

ID: 224961632