ArticleMicrobial biotechnology2022
Highly efficient rDNA-mediated multicopy integration based on the dynamic balance of rDNA in Saccharomyces cerevisiae.
Article in Microbial biotechnology, 2022. 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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11 citing papers in PubMed, 21 citations in OpenAlex.
- Fungal expression strategies for heterologous production of antimicrobial peptides.Fungal biology and biotechnology · 2026Review
- rDNA-mediated multicopy integration and gene dosage quantification system for microbial zeaxanthin biosynthesis.World journal of microbiology & biotechnology · 2026Article
- CRISPR-based synthetic biology toolkit development inSynthetic and systems biotechnology · 2026Article
- Efficient Introduction of Genetic Elements at Specific Insertion Loci.Methods in molecular biology (Clifton, N.J.) · 2026Article
- Optimizing multicopy chromosomal integration for stable high-performing strains.Nature chemical biology · 2024Article
- A Novel Biosynthetic Strategy for Ginsenoside Ro: Construction of a Metabolically EngineeredInternational journal of molecular sciences · 2024Article
- Enhanced heterologous gene expression in Trichoderma reesei by promoting multicopy integration.Applied microbiology and biotechnology · 2024Article
- Customizable and stable multilocus chromosomal integration: a novel glucose-dependent selection system in Aureobasidium spp.Biotechnology for biofuels and bioproducts · 2024Article
- EngineeringEngineering microbiology · 2024Review
- Highly efficient rDNA-mediated multicopy integration based on the dynamic balance of rDNA in Saccharomyces cerevisiae.Microbial biotechnology · 2022Article
- Review
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Authors and funding
8 authors at 1 institution in 1 country.
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No grant is acknowledged in the PubMed record.
Abstract
Engineered Saccharomyces cerevisiae strains are good cell factories, and developing additional genetic manipulation tools will accelerate construction of metabolically engineered strains. Highly repetitive rDNA sequence is one of two main sites typically used for multicopy integration of genes. Here, we developed a simple and high-efficiency strategy for rDNA-mediated multicopy gene integration based on the dynamic balance of rDNA in S. cerevisiae. rDNA copy number was decreased by pre-treatment with hydroxyurea (HU). Then, heterologous genes were integrated into the rDNA sequence. The copy number of the integrated heterologous genes increased along with restoration of the copy number of rDNA. Our results demonstrated that HU pre-treatment doubled the number of integrated gene copies; moreover, compared with removing HU stress during transformation, removing HU stress after selection of transformants had a higher probability of resulting in transformants with high-copy integrated genes. Finally, we integrated 18.0 copies of the xylose isomerase gene into the S. cerevisiae genome in a single step. This novel rDNA-mediated multicopy genome integration strategy provides a convenient and efficient tool for further metabolic engineering of S. cerevisiae.
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