Journal
ACS SYNTHETIC BIOLOGY
Volume 3, Issue 3, Pages 188-191Publisher
AMER CHEMICAL SOC
DOI: 10.1021/sb400091p
Keywords
synthetic promoters; core promoter construction; promoter library; transcriptional fine-tuning; Pichia pastoris
Categories
Funding
- Austrian Science Fund (FWF) [W901]
- FFG [824186]
- bmvit
- mvwfi
- ZIT
- Zukunftsstiftung Tirol and Land Steiermark
- Austrian Science Fund (FWF) [W 901] Funding Source: researchfish
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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.
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