Saccharomyces cerevisiaesingle-copy plasmids for auxotrophy compensation, multiple marker selection, and for designing metabolically cooperating communities.
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 typicalSaccharomyces cerevisiaelaboratory strains. The 23 single-copy plasmids complement for deficiencies inHIS3, LEU2, URA3, MET17 or LYS2genes 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.
Department of Biochemistry and Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.|Mill Hill Laboratory, The Francis Crick Institute, London, UK.
Michael
M
Mülleder
Department of Biochemistry and Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.|Chalmers University of Technology, Gothenburg, Sweden.
Kate
K
Campbell
Department of Biochemistry and Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.
Olga
O
Matsarskaia
Department of Biochemistry and Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.
Florian
F
Eckerstorfer
Department of Biochemistry and Cambridge Systems Biology Centre, University of Cambridge, Cambridge, UK.|Mill Hill Laboratory, The Francis Crick Institute, London, UK.
Markus
M
Ralser
Saccharomyces cerevisiae; auxotrophic markers; centromeric plasmid; metabolism; self-establishing metabolically cooperating (SeMeCo) communities