Use of Yeast Plasmids: Transformation and Inheritance Assays

Abstract

The use of the budding yeast Saccharomyces cerevisiae as a model genetic organism has been facilitated by the availability of a wide range of yeast shuttle vectors, plasmids that can be propagated in Escherichia coli and also in yeast, where they are stably maintained at low- or high-copy number, depending on the plasmid system. Here we provide an introduction to the low-copy (ARS/CEN) and multi-copy (2-μm-based) plasmids, the marker genes commonly used for plasmid selection in yeast, methods for transforming yeast and monitoring plasmid inheritance, and tips for working with yeast transformants.

Keywords: 2-μm plasmid; ARS/CEN plasmid; Autonomously replicating sequence (ARS); Centromere (CEN); Partitioning; Plasmid copy number; Plating assays; Selectable marker genes; Yeast transformation.