Saccharomyces cerevisiae single-copy plasmids for auxotrophy compensation, multiple marker selection, and for designing metabolically cooperating communities
Journal article, 2016

Auxotrophic markers are useful tools in cloning and genome editing, enable a large spectrum of genetic techniques, as well as facilitate the study of metabolite exchange interactions in microbial communities. If unused background auxotrophies are left uncomplemented however, yeast cells need to be grown in nutrient supplemented or rich growth media compositions, which precludes the analysis of biosynthetic metabolism, and which leads to a profound impact on physiology and gene expression. Here we present a series of 23 centromeric plasmids designed to restore prototrophy in typical Saccharomyces cerevisiae laboratory strains. The 23 single-copy plasmids complement for deficiencies in HIS3, LEU2, URA3, MET17 or LYS2 genes and in their combinations, to match the auxotrophic background of the popular functional-genomic yeast libraries that are based on the S288c strain. The plasmids are further suitable for designing self-establishing metabolically cooperating (SeMeCo) communities, and possess a uniform multiple cloning site to exploit multiple parallel selection markers in protein expression experiments.

Saccharomyces cerevisiae

Auxotrophic markers

Self-establishing metabolically cooperating (SeMeCo) communities

Centromeric plasmid

Metabolism

Author

Michael Mülleder

The Francis Crick Institute

University of Cambridge

Kate Campbell

Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology

University of Cambridge

Olga Matsarskaia

University of Cambridge

Florian Eckerstorfer

University of Cambridge

Markus Ralser

The Francis Crick Institute

University of Cambridge

F1000Research

2046-1402 (ISSN)

Vol. 5 2351

Areas of Advance

Health Engineering

Subject Categories

Microbiology

Bioinformatics and Systems Biology

Genetics

DOI

10.12688/f1000research.9606.1

More information

Latest update

11/12/2020