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Heterologous Expression in Budding Yeast as a Tool for Studying the Plant Cell Morphogenesis Machinery

  • Fatima Cvrčková
  • Michal Hála
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1080)

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.

Key words

Saccharomyces cerevisiae Heterologous gene expression Inducible expression system Complementation Phenotype rescue cDNA library screening 

Notes

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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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Fatima Cvrčková
    • 1
  • Michal Hála
    • 2
  1. 1.Department of Experimental Plant Biology, Faculty of ScienceCharles UniversityPragueCzech Republic
  2. 2.Department of Experimental Plant Biology, Faculty of ScienceCharles University and Institute of Experimental Botany, Academy of Sciences of the Czech RepublicPragueCzech Republic

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