Auxotrophy-Independent Plasmid Shuttle Vectors for Applications in Diverse Yeasts

Author:

Smith Jeremy1ORCID,Sislak Christine1,Fernandez Mendoza Pedro1,Carmichael Laurin1,Lewis Alisha1,Chen Anqi12ORCID,Jiang Glycine1,Gibney Patrick1ORCID

Affiliation:

1. Department of Food Science, Cornell University, Ithaca, NY 14853, USA

2. Science Center for Future Foods, Jiangnan University, Wuxi 214122, China

Abstract

Plasmid shuttle vectors are a common tool used to study yeast physiology. The majority of yeast plasmids have been optimized for Saccharomyces cerevisiae lab strain compatibility, relying on auxotrophic complementation as their selective property. We sought to construct a series of plasmid shuttle vectors to extend functionality beyond strains with auxotrophic requirements, and test compatibility across a diverse panel of yeasts. We constructed 18 plasmids which were successfully maintained by yeasts from several genera. From a panel of 24 yeast strains, these plasmids were maintained by 18 yeasts, spanning 11 species within the genera Lachancea, Metschnikowia, Pichia, Saccharomyces, and Torulaspora. Additionally, an integrated gene expression reporter was assayed for functional compatibility with the 18 strains. Plasmid-derived gene expression was observed for 13 strains, spanning five species within the Saccharomyces genus, in addition to Torulaspora delbrueckii. These results indicate that this plasmid series is broadly useful for advancements and applications within academia, biotechnology, and the food and fermentation industries for research utilizing diverse Saccharomyces and non-Saccharomyces yeasts.

Funder

U.S. Department of Agriculture, National Institute of Food and Agriculture, Multistate Hatch Federal Capacity Funds

Publisher

MDPI AG

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