ArticleApplied and environmental microbiology2006
Engineering of promoter replacement cassettes for fine-tuning of gene expression in Saccharomyces cerevisiae.
Article in Applied and environmental microbiology, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 90 papers.
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90 citing papers in PubMed, 225 citations in OpenAlex.
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- A Standardized Set of MoClo-Compatible Inducible Promoter Systems for Tunable Gene Expression in Yeast.ACS synthetic biology · 2024Article
- Systematic sequence engineering enhances the induction strength of the glucose-regulated GTH1 promoter of Komagataella phaffii.Nucleic acids research · 2023Article
- Design of synthetic promoters for cyanobacteria with generative deep-learning model.Nucleic acids research · 2023Article
- Current genetic strategies to investigate gene functions in Trichoderma reesei.Microbial cell factories · 2023Review
- Directed evolution of a wax ester synthase for production of fatty acid ethyl esters in Saccharomyces cerevisiae.Applied microbiology and biotechnology · 2023Article
- Design of Four Small-Molecule-Inducible Systems in the Yeast Chromosome, Applied to Optimize Terpene Biosynthesis.ACS synthetic biology · 2023Article
- Natural promoters and promoter engineering strategies for metabolic regulation in Saccharomyces cerevisiae.Journal of industrial microbiology & biotechnology · 2023Review
- Transcription regulation strategies in methylotrophs: progress and challenges.Bioresources and bioprocessing · 2022Review
- Identification of acetic acid sensitive strains through biosensor-based screening of a Saccharomyces cerevisiae CRISPRi library.Microbial cell factories · 2022Article
- Identification of the Talaromyces cellulolyticus Gene Encoding an Extracellular Enzyme with β-galactosidase Activity and Testing it as a Reporter for Gene Expression Assays.Molecular biotechnology · 2022Article
- Expanding the promoter toolbox for metabolic engineering of methylotrophic yeasts.Applied microbiology and biotechnology · 2022Review
- Characterizing and engineering promoters for metabolic engineering ofSynthetic and systems biotechnology · 2022Article
30 more citing papers are in PubMed but not listed here.
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The strong overexpression or complete deletion of a gene gives only limited information about its control over a certain phenotype or pathway. Gene function studies based on these methods are therefore incomplete. To effect facile manipulation of gene expression across a full continuum of possible expression levels, we recently created a library of mutant promoters. Here, we provide the detailed characterization of our yeast promoter collection comprising 11 mutants of the strong constitutive Saccharomyces cerevisiae TEF1 promoter. The activities of the mutant promoters range between about 8% and 120% of the activity of the unmutated TEF1 promoter. The differences in reporter gene expression in the 11 mutants were independent of the carbon source used, and real-time PCR confirmed that these differences were due to varying levels of transcription (i.e., caused by varying promoter strengths). In addition to a CEN/ARS plasmid-based promoter collection, we also created promoter replacement cassettes. They enable genomic integration of our mutant promoter collection upstream of any given yeast gene, allowing detailed genotype-phenotype characterizations. To illustrate the utility of the method, the GPD1 promoter of S. cerevisiae was replaced by five TEF1 promoter mutants of different strengths, which allowed analysis of the impact of glycerol 3-phosphate dehydrogenase activity on the glycerol yield.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.