title: Synthetic core promoters for Pichia pastoris.
abstract: Synthetic promoters are commonly used tools for circuit design or high level protein production. Promoter engineering efforts in yeasts, such as Saccharomyces cerevisiae and Pichia pastoris have mostly been focused on altering upstream regulatory sequences such as transcription factor binding sites. In higher eukaryotes synthetic core promoters, directly needed for transcription initiation by RNA Polymerase II, have been successfully designed. Here we report the first synthetic yeast core promoter for P. pastoris, based on natural yeast core promoters. Furthermore we used this synthetic core promoter sequence to engineer the core promoter of the natural AOX1 promoter, thereby creating a set of core promoters providing a range of different expression levels. As opposed to engineering strategies of the significantly longer entire promoter, such short core promoters can directly be added on a PCR primer facilitating library generation and are sufficient to obtain variable expression yields.
authors.affiliation: Institute of Molecular Biotechnology, Graz University of Technology , Petersgasse 14, Graz A-8010, Austria.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vogl
authors.forename: Claudia
authors.initials: C
authors.lastname: Ruth
authors.forename: Julia
authors.initials: J
authors.lastname: Pitzer
authors.forename: Thomas
authors.initials: T
authors.lastname: Kickenweiz
authors.forename: Anton
authors.initials: A
authors.lastname: Glieder
mesh_terms: D001483:Base Sequence; D005800:Genes, Fungal / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D016415:Sequence Alignment; D058615:Synthetic Biology / Q000379:methods*