Expression vectors were created in which the 5′ end of the Saccharomyces cerevisiae CDC9 gene, which encodes a mitochondrial targeting peptide, was cloned in-frame with the coding regions of the EcoR I, Hind III, and Pst I endonuclease genes. Expression of the EcoR I and Hind III fusion proteins inhibited growth of yeast on glycerol-containing media and resulted in the nearly quantitative restriction digestion of their mitochondrial DNA. In contrast, expression of Pst I, which does not recognize any sites within yeast mitochondrial DNA, had no effect on growth in glycerol-containing media, and did not affect the integrity of the mitochondrial genome.
Bibliographical noteFunding Information:
We thank Drs Barbara Garvik and Lee Hartwell as well as Brendan Cormack, Jasper Rine, and Masimo Zeviani for kindly providing yeast strains and reagents used herein. We also thank Dr Modrich for kindly providing Eco R I antibody. This work was supported in part by a grant from the NIH (AG16678) and by a grant from the American Heart Association (AHA MN 9951198Z).
- Restriction endonuclease
- Saccharomyces cerevisiae