Heterologous Expression in Budding Yeast as a Tool for Studying the Plant Cell Morphogenesis Machinery - Plant Cell Morphogenesis: Methods and Protocols - page 256

Chapter 23

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

Fatima Cvr ˇ ková and Michal Hála

Abstract

The budding yeast ( Saccharomyces cerevisiae ) can serve as a unique experimental system for functional stud-
ies 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 sum-
marize 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

1 Introduction

After decades as a versatile model system of its own merit, the bud-
ding yeast Saccharomyces cerevisiae has become fi rmly established
also in laboratories focusing on other eukaryotes, though mainly
only as a “living test-tube” utilized for the detection of protein-
protein interactions in various modifi cations of the yeast two-
hybrid system, or Y2H [ 1 ]. While the Y2H itself is out of scope of
this chapter, it helped to introduce routine work with yeast into
many laboratories, making the use of additional yeast-based experi-
mental strategies easier.
Evolutionarily conserved components of the cellular machineries—
such as the cytoskeleton or the membrane-traffi cking system—
provide the most promising targets for yeast-based functional
experiments. Several experimental strategies can utilize the knowl-
edge gained in yeast to study plant cell morphogenesis; specifi c
examples are provided in the references cited below.