In vivo analysis of mtDNA replication defects in yeast

Methods. 2010 Aug;51(4):426-36. doi: 10.1016/j.ymeth.2010.02.023. Epub 2010 Mar 2.

Abstract

The yeast Saccharomyces cerevisiae has the capacity to survive large deletions or total loss of mtDNA (petite mutants), and thus in the last few years it has been used as a model system to study defects in mitochondrial DNA (mtDNA) maintenance produced by mutations in genes involved in mtDNA replication. In this paper we describe methods to obtain strains harboring mutations in nuclear genes essential for the integrity of mtDNA, to measure the frequency and the nature of petite mutants, to estimate the point mutation frequency in mtDNA and to determine whether a nuclear mutation is recessive or dominant and, in the latter case, the kind of dominance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • DNA Primers / genetics
  • DNA Replication / genetics*
  • DNA, Fungal / biosynthesis*
  • DNA, Fungal / genetics*
  • DNA, Mitochondrial / biosynthesis*
  • DNA, Mitochondrial / genetics*
  • Fluorescent Dyes
  • Genes, Fungal
  • Genetic Techniques
  • Indoles
  • Mutagenesis
  • Mutation*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Staining and Labeling

Substances

  • DNA Primers
  • DNA, Fungal
  • DNA, Mitochondrial
  • Fluorescent Dyes
  • Indoles
  • DAPI