This chapter examines the yeast mitochondrial genes. The mitochondrial genome of Saccharomyces cerevisiae is a circular duplex molecule consisting of 70–75 kbp. Two different approaches have been used to sequence yeast mitochondrial genes. The transfer RNA (tRNA) and ribosomal RNA genes are sequenced from fragments of mitochondrial DNA (mtDNA) cloned in bacteria. Most of the sequences of the protein coding genes are obtained from the genomes of cytoplasmic petite (p-) mutants. The procedures for the isolation of genetically marked p- mutants and their use in the analysis of mitochondrial genes are described in this chapter. p- mutants are selected for the retention of known genetic markers in mtDNA. The p- clones are purified and checked for the stability of their genotypes. The physical locations of the p- genomes on the wild-type map can usually be established by restriction mapping. Clones with coherent segments of mtDNA free of internal deletions or rearrangements are used for sequence analysis.
ASJC Scopus subject areas
- Molecular Biology