The budding yeast Saccharomyces cerevisiae is a widely utilized model system with myriad applications in terms of industrial, biotechnology, and synthetic biology purposes. One such application is the biosynthesis of commercially and medically important bioactive compounds and their precursors, which oftentimes require culturing conditions at low temperatures to optimize production yield rather than cellular fitness. To lend insight into genetic modifications that may assist this goal, this work focuses on a systematic analysis of the genes that result in an increase in survival following freezing. At present, these genes have been identified in a wide variety of S. cerevisiae wild-type backgrounds-that vary significantly in their properties and behaviors-and in the conditions that led to the annotation of the freeze-thaw survival phenotype. In this work, we report a complete characterization of the thermal tolerance and viability for the freeze-thaw gene family following a standardized protocol within a unified genetic background, the extensively used BY4741 laboratory strain. Our results reveal that five out of these six genes are linked to increased viability in response to both freeze-thaw stress as well as enhanced survival during a heat shock stressor. Follow-up analysis characterized the local spatial effects that gene modification at each locus causes when utilizing the common kanamycin resistance cassette (KanMX6) for the creation of mutant strains and engineering purposes.

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Journal Article

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Hartnett D, Dotto M, Aguirre A, Brandao S, Chauca M, Chiang S, Cronin M, Deokar N, Martin A, McCune Z, ... Show all

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