title: The development and characterization of synthetic minimal yeast promoters.
abstract: Synthetic promoters, especially minimally sized, are critical for advancing fungal synthetic biology. Fungal promoters often span hundreds of base pairs, nearly ten times the amount of bacterial counterparts. This size limits large-scale synthetic biology efforts in yeasts. Here we address this shortcoming by establishing a methodical workflow necessary to identify robust minimal core elements that can be linked with minimal upstream activating sequences to develop short, yet strong yeast promoters. Through a series of library-based synthesis, analysis and robustness tests, we create a set of non-homologous, purely synthetic, minimal promoters for yeast. These promoters are comprised of short core elements that are generic and interoperable and 10 bp UAS elements that impart strong, constitutive function. Through this methodology, we are able to generate the shortest fungal promoters to date, which can achieve high levels of both inducible and constitutive expression with up to an 80% reduction in size.
authors.affiliation: Department of Molecular Biosciences, The University of Texas at Austin, 2500 Speedway Avenue, Austin, Texas 78712, USA.
authors.forename: Heidi
authors.initials: H
authors.lastname: Redden
authors.affiliation: 1] Department of Molecular Biosciences, The University of Texas at Austin, 2500 Speedway Avenue, Austin, Texas 78712, USA. [2] Department of Chemical Engineering, The University of Texas at Austin, 200 E Dean Keeton St. Stop C0400, Austin, Texas 78712, USA.
authors.forename: Hal S
authors.initials: HS
authors.lastname: Alper
mesh_terms: D004271:DNA, Fungal / Q000235:genetics*; D004926:Escherichia coli; D005434:Flow Cytometry; D015723:Gene Library; D005813:Genes, Synthetic; D005818:Genetic Engineering / Q000379:methods*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D058615:Synthetic Biology / Q000379:methods; D057188:Workflow
chemical_list: D004271:DNA, Fungal