ArticleBioMed research international2013
Combinatorial mutagenesis and selection to understand and improve yeast promoters.
Article in BioMed research international, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed, 31 citations in OpenAlex.
- Next-generation stress-inducible Komagataella phaffii promoter variants.Microbial cell factories · 2025Article
- Functional consequences of modification of the photosystem I/photosystem II ratio in the cyanobacteriumJournal of bacteriology · 2024Article
- Systematic sequence engineering enhances the induction strength of the glucose-regulated GTH1 promoter of Komagataella phaffii.Nucleic acids research · 2023Article
- Engineering of Promoters for Gene Expression in Pichia pastoris.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Engineering yeast artificial core promoter with designated base motifs.Microbial cell factories · 2020Article
- Construction and characterization of centromeric plasmids for Komagataella phaffii using a color-based plasmid stability assay.PloS one · 2020Article
- Synthetic Core Promoters as Universal Parts for Fine-Tuning Expression in Different Yeast Species.ACS synthetic biology · 2017Article
- Recent Progress on Systems and Synthetic Biology Approaches to Engineer Fungi As Microbial Cell Factories.Current genomics · 2016Article
- Synthetic core promoters for Pichia pastoris.ACS synthetic biology · 2014Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Microbial promoters are important targets both for understanding the global gene expression and developing genetic tools for heterologous expression of proteins and complex biosynthetic pathways. Previously, we have developed and used combinatorial mutagenesis methods to analyse and improve bacterial expression systems. Here, we present for the first time an analogous strategy for yeast. Our model promoter is the strong and inducible P AOX1 promoter in methylotrophic Pichia pastoris. The Zeocin resistance gene was applied as a valuable reporter for mutant P AOX1 promoter activity, and we used an episomal plasmid vector to ensure a constant reporter gene dosage in the yeast host cells. This novel design enabled direct selection for colonies of recombinant cells with altered Zeocin tolerance levels originating solely from randomly introduced point mutations in the P AOX1 promoter DNA sequence. We demonstrate that this approach can be used to select for P AOX1 promoter variants with abolished glucose repression in large mutant libraries. We also selected P AOX1 promoter variants with elevated expression level under induced conditions. The properties of the selected P AOX1 promoter variants were confirmed by expressing luciferase as an alternative reporter gene. The tools developed here should be useful for effective screening, characterization, and improvement of any yeast promoters.
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