Saccharomyces cerevisiae Shuttle vectors

Abstract

Yeast shuttle vectors are indispensable tools in yeast research. They enable cloning of defined DNA sequences in Escherichia coli and their direct transfer into Saccharomyces cerevisiae cells. There are three types of commonly used yeast shuttle vectors: centromeric plasmids, episomal plasmids and integrating plasmids. In this review, we discuss the different plasmid systems and their characteristic features. We focus on their segregational stability and copy number and indicate how to modify these properties. Copyright © 2017 John Wiley & Sons, Ltd.

The art of vector engineering: towards the construction of next‐generation genetic tools

“…The YEps are based on the endogenous 2μ plasmid mentioned above with addition of a bacterial origin of replication and selection marker, yeast selection marker and the expression cassette. Still, the YIps need to have homology sequences so they can integrate in the yeast genome via homologous recombination (Gnügge and Rudolf, ). For all of them, the selection marker is usually auxotrophic, which somewhat restricts their use because there is usually only URA3 (encoding orotidine‐5′‐phosphate decarboxylase), LEU2 (encoding 3‐isopropylmalate dehydrogenase), HIS3 (encoding imidazoleglycerol‐phosphate dehydratase) and TRP1 (encoding phosphoribosylanthranilate isomerase) options, besides the need for a strain with the original gene deleted.…”

“…Table summarizes the most important tools for yeast manipulation from 2007 to 2017. Likewise, Gnügge and Rudolf () also described, in an extended review, most of the shuttle vectors available for yeast.…”

“…Another crucial component for a vector is the selection marker, which can be any gene allowing a selective advantage to the positive transformants, ranging from auxotrophy (corresponding to a metabolic enzyme missing in the host genome) to drug resistance (Gnügge and Rudolf, ). Also, a multiple cloning site (MCS) is usually added to facilitate cloning of the desired DNA, containing several sites recognized by different restriction enzymes.…”

YHp as a highly stable, hyper‐copy, hyper‐expression plasmid constructed using a full 2‐μm circle sequence in cir0 strains of Saccharomyces cerevisiae

“…Various vector systems have been developed for gene manipulation in the yeast Saccharomyces cerevisiae , depending on the purpose of the studies. Some examples are yeast integrating plasmid (YIp), yeast replicating plasmid (YRp), yeast centromeric plasmid (YCp), and YEp (Gnugge & Rudolf, ; Gunge, ). YIp is a chromosomal integrative vector possessing only selection marker gene.…”

Development and Optimisation of a Defined High Cell Density Yeast Medium

“…In contrast to many other model organisms, yeasts offer comparatively simple genetic manipulation using either plasmid-based expression or genome modifications [11]. Over the last 70 years, the community established a series of commonly used selection markers for this purpose, but their use requires a defined growth media for auxotrophic selection [12]. Most commonly, a defined growth media uses Glucose as its only carbon source with concentrations varying between 10 g/ up to 60 g/ .…”