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Expression of Barstar as a selectable marker in yeast mitochondria

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Abstract

We describe a new and potentially universal selection system for mitochondrial transformation based on bacterial genes, and demonstrate its feasibility in Saccharomyces cerevisiae. We first found that cytoplasmically synthesized Barnase, an RNase, interferes with mitochondrial gene expression when targeted to the organelle, without causing lethality when expressed at appropriate levels. Next, we synthesized a gene that uses the yeast mitochondrial genetic code to direct the synthesis of the specific Barnase inhibitor Barstar, and demonstrated that expression of this gene, BARSTM, integrated in mtDNA protects respiratory function from imported barnase. Finally, we showed that screening for resistance to mitochondrially targeted barnase can be used to identify rare mitochondrial transformants that had incorporated BARSTM in their mitochondrial DNA. The possibility of employing this strategy in other organisms is discussed.

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Acknowledgements

This work was supported by the Institut National de la Recherche Agronomique (INRA, France) and a grant from US National Institutes of Health (GM29362). We thank N. Bonnefoy for helpful discussions and technical help.

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Authors and Affiliations

  1. Station de Génétique et d'Amélioration des Plantes, INRA, Route de Saint-Cyr, 78026, Versailles, France

    H. Mireau & N. Arnal

  2. Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853-2703, USA

    T. D. Fox

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  1. H. Mireau
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  2. N. Arnal
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  3. T. D. Fox
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Correspondence to H. Mireau.

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Communicated by R. G. Herrmann

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Mireau, H., Arnal, N. & Fox, T.D. Expression of Barstar as a selectable marker in yeast mitochondria. Mol Gen Genomics 270, 1–8 (2003). https://doi.org/10.1007/s00438-003-0879-2

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  • DOI: https://doi.org/10.1007/s00438-003-0879-2

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