Genetic Transformation of Saccharomyces cerevisiae and Chlamydomonas reinhardtii Mitochondria

Abstract

Publisher Summary This chapter focuses largely on the more highly developed saccharomyces cerevisiae system to summarize some basic features of yeast mitochondrial genetics, describes the current methods for the delivery of DNA into the organelle, and outlines the strategies to create directed mutations in mitochondrial genes and insert new genes into mitochondrial DNA (mtDNA). It also presents the general strategy and protocol for chlamydomonas mitochondrial transformation. The methods developed for manipulation of the saccharomyces cerevisiae mitochondrial genome have already been useful to set up an efficient technique for delivery of mtDNA into chlamydomonas mitochondria and should also provide a useful model for other systems such as the petite-negative yeast S. pombe. So far, transformation of chlamydomonas requires the selection of respiratory proficiency thus allowing the creation of mutation in the nd genes, but not in cob or cox1. Although limited, this technique is of importance because this opens, for the first time, the way to in vitro mutagenesis of organelle genes for complex I, because saccharomyces cerevisiae does not contain a bona fide complex I. As appropriate selectable markers are developed for chlamydomonas and other species, it seems likely that their full mitochondrial genomes will become amenable to in vivo experimental analysis, providing alternative model systems to saccharomyces cerevisiae.