Developing methods and strains for genetic studies in the Saccharomyces bayanus var. uvarum species

Abstract

For years, Saccharomyces cerevisiae has been used as a model organism to gain insight into complex biological processes. The study of closely related yeast species may be critical for understanding the molecular mechanism of evolution. Among those species, S. bayanus var. uvarum could be particularly pertinent because of the availability of its genome sequence. However, to date, in that species genetic studies are problematical due to the lack of standard strains collection and genetic methods. Here, we have developed heterothallic S. bayanus var. uvarum strains and obtained stable haploid strains. We further used UV-induced mutation and gene disruption to create a collection of auxotrophic derivatives. Finally, we have elaborated or improved methods to cultivate cells, obtain zygotes and spores and to transform this species. All these tools can now be used by the scientific community to study the biology of this species.

Evolution of gene network activity by tuning the strength of negative-feedback regulation

“…The resulting plasmid was integrated into the P GAL1 locus of the wild-type S. paradoxus strain by using a previously described 30 transformation technique. The complete sequences of the GAL network promoters and genes from both yeast species are shown in Supplementary Methods.…”

The [URE3] Prion Is Not Conserved Among Saccharomyces Species

“…Details of the constructions are available upon request. S. uvarum Su [ure0] was constructed from strain Su5-1A (Talarek et al 2004) by transformation with a PCR product containing the dal5 Su ::ADE2 Sc cassette amplified from pYe2L-pDAL5 Su ::ADE2 Sc (the correct notation should be dal5::P DAL5 ADE2; for the sake of simplicity we noted the construction as pDAL5::ADE2). Integrative transformants were selected on minimal medium without adenine and with proline as poor nitrogen source.…”

“…All mating, sporulation, and dissection procedures were carried out according to standard protocols (Sherman and Hicks 1991). For S. uvarum and S. paradoxus species, procedures were performed as previously described (Talarek et al 2004). Asci dissections were performed by micromanipulation (Singer Instrument MSM).…”

Natural Populations of Saccharomyces kudriavzevii in Portugal Are Associated with Oak Bark and Are Sympatric with S. cerevisiae and S. paradoxus

“…They are used as fermenting agents worldwide and stand out as model eukaryotic organisms in various fields of biological science ranging from biochemistry to genomics. The modern phylogenetic concept of Saccharomyces restricts this genus to eight species: S. bayanus, S. cerevisiae, S. paradoxus, S. pastorianus, S. uvarum, S. cariocanus, S. kudriavzevii, and S. mikatae (14,24,28,33). Most of these species are delimited by postzygotic isolation, but S. pastorianus and S. bayanus are thought to be hybrid species, since their genomes have contributions from both S. cerevisiae and S. uvarum (formerly designated S. bayanus var.…”