ArticleCurrent genetics2007
Use of MET3 promoters for regulated gene expression in Ashbya gossypii.
Article in Current genetics, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 29 citations in OpenAlex.
- Use of the Saccharomycopsis schoenii MET17 promoter for regulated heterologous gene expression.Current genetics · 2024Article
- New Promoters for Metabolic Engineering ofJournal of fungi (Basel, Switzerland) · 2021Article
- Sporulation inJournal of fungi (Basel, Switzerland) · 2020Review
- Overexpression ofMicroorganisms · 2019Article
- Toxicity of Potential Fungal Defense Proteins towards the Fungivorous Nematodes Aphelenchus avenae and Bursaphelenchus okinawaensis.Applied and environmental microbiology · 2018Article
- The Promoter Toolbox for Recombinant Gene Expression inFrontiers in bioengineering and biotechnology · 2018Review
- L-Methionine repressible promoters for tuneable gene expression in Trichoderma reesei.Microbial cell factories · 2015Article
- Increased production of inosine and guanosine by means of metabolic engineering of the purine pathway in Ashbya gossypii.Microbial cell factories · 2015Article
- Repressible promoters - a novel tool to generate conditional mutants in Pichia pastoris.Microbial cell factories · 2013Article
- Combination of ERG9 Repression and Enzyme Fusion Technology for Improved Production of Amorphadiene in Saccharomyces cerevisiae.Journal of analytical methods in chemistry · 2013Article
- N-acetylglucosamine utilization by Saccharomyces cerevisiae based on expression of Candida albicans NAG genes.Applied and environmental microbiology · 2009Article
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Authors and funding
2 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A central tool for gene function analysis is the construction mutant strains. This can be done conveniently in A. gossypii using PCR-based tools. The deletion of essential genes can be performed since initial transformants are sheltered in a heterokaryotic mycelium, which contains nuclei with both wild type and mutant alleles. The analysis of mutant phenotypes in A. gossypii is regularly started by germinating spores, which contain only one nucleus. Thus, selection can be used to identify mutant germ cells and germlings. However, such an analysis yields only mutant mycelia if the deleted gene is not essential. We describe the use of the regulatable Saccharomyces cerevisiae and A. gossypii MET3 promoters as novel tools to regulate gene expression in A. gossypii. Conditional expression was tested using GFP and lacZ-reporter genes. Regulation of MET3 promoters was found to be dependent on methionine but not on cysteine and down-regulation to about 1/10 of the initial expression levels was achieved. We used the A. gossypii WAL1 and CYK1 genes as models to demonstrate that MET3 promoters could regulate the expression of these genes and reveal their mutant phenotypes depending on the presence or absence of methionine. Finally, we show that the AgMET3 promoter contains two Cpf1-binding sites and that AgCPF1 can complement the S. cerevisiae cpf1 methionine auxotrophy.
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