Abstract
The budding yeast (Saccharomyces cerevisiae) can serve as a unique experimental system for functional studies of heterologous genes, allowing not only complementation of readily available yeast mutations but also generation of overexpression phenotypes and in some cases also rescue of such phenotypes. Here we summarize the main considerations that have to be taken into account when using the yeast expression system for investigating the function of plant genes participating in cell morphogenesis; outline the strategies of experiment planning, yeast strain selection (or construction), and expression vector choice; and provide detailed protocols for yeast transformation, transformant selection, and phenotype evaluation.
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Acknowledgments
This work has been supported by the MSM0021620858 project. We thank Marta Čadyová for expert technical assistance and members of the A. Ragnini laboratory (University of Vienna, Austria) for the rapid yeast transformation protocol.
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Cvrčková, F., Hála, M. (2014). Heterologous Expression in Budding Yeast as a Tool for Studying the Plant Cell Morphogenesis Machinery. In: Žárský, V., Cvrčková, F. (eds) Plant Cell Morphogenesis. Methods in Molecular Biology, vol 1080. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-643-6_23
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DOI: https://doi.org/10.1007/978-1-62703-643-6_23
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Publisher Name: Humana Press, Totowa, NJ
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Online ISBN: 978-1-62703-643-6
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