Pichia pastoris Alcohol Oxidase 1 (AOX1) Core Promoter Engineering by High Resolution Systematic Mutagenesis

Pichia pastoris Alcohol Oxidase 1 (AOX1) Core Promoter Engineering by High Resolution Systematic... IntroductionEukaryotic promoters can generally be separated in two parts: an upstream regulatory sequence (URS, also termed cis‐regulatory modules, CRMs, or enhancers) and a core promoter. CRMs contain transcription factor binding sites (TFBSs), that are bound by specific transcription factors (TFs), conferring, for example, cell‐cycle or carbon source dependent regulation. In contrast, the core promoter typically controls transcription initiation, as RNA polymerase II (RNAPII) and general TFs bind to this region. Gaining insights on core promoter sequence‐function relationship is essential for understanding transcription initiation and for generating core promoters variants to increase or fine‐tune expression for synthetic biology and metabolic engineering applications and for the design of general expression vectors.In higher eukaryotes, synthetic core promoters have been designed based on commonly occurring motifs such as TATA box, Inr (initator), DPE (downstream core promoter element), and MTE (motif ten element. In lower eukaryotes, namely yeasts, the only clearly conserved motif in core promoters appears to be the TATA box. Several studies have demonstrated the feasibility of synthetic core promoter design in yeasts. Different methodologies were applied for promoter designs in Saccharomyces cerevisiae and Komagataella phaffii (syn. Pichia pastoris), namely random mutagenesis, indels (insertions and deletions) in the 5′ untranslated region (5′UTR) http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology Journal Wiley

Pichia pastoris Alcohol Oxidase 1 (AOX1) Core Promoter Engineering by High Resolution Systematic Mutagenesis

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Publisher
Wiley Subscription Services, Inc., A Wiley Company
Copyright
© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
ISSN
1860-6768
eISSN
1860-7314
D.O.I.
10.1002/biot.201700340
Publisher site
See Article on Publisher Site

Abstract

IntroductionEukaryotic promoters can generally be separated in two parts: an upstream regulatory sequence (URS, also termed cis‐regulatory modules, CRMs, or enhancers) and a core promoter. CRMs contain transcription factor binding sites (TFBSs), that are bound by specific transcription factors (TFs), conferring, for example, cell‐cycle or carbon source dependent regulation. In contrast, the core promoter typically controls transcription initiation, as RNA polymerase II (RNAPII) and general TFs bind to this region. Gaining insights on core promoter sequence‐function relationship is essential for understanding transcription initiation and for generating core promoters variants to increase or fine‐tune expression for synthetic biology and metabolic engineering applications and for the design of general expression vectors.In higher eukaryotes, synthetic core promoters have been designed based on commonly occurring motifs such as TATA box, Inr (initator), DPE (downstream core promoter element), and MTE (motif ten element. In lower eukaryotes, namely yeasts, the only clearly conserved motif in core promoters appears to be the TATA box. Several studies have demonstrated the feasibility of synthetic core promoter design in yeasts. Different methodologies were applied for promoter designs in Saccharomyces cerevisiae and Komagataella phaffii (syn. Pichia pastoris), namely random mutagenesis, indels (insertions and deletions) in the 5′ untranslated region (5′UTR)

Journal

Biotechnology JournalWiley

Published: Jan 1, 2018

Keywords: ; ; ; ;

References

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