ArticleApplied and environmental microbiology2009
Phleomycin increases transformation efficiency and promotes single integrations in Schizophyllum commune.
Article in Applied and environmental microbiology, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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26 citing papers in PubMed, 61 citations in OpenAlex.
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- Inositol Signaling in the Basidiomycete FungusJournal of fungi (Basel, Switzerland) · 2021Article
- H3K4me2 ChIP-Seq reveals the epigenetic landscape during mushroom formation and novel developmental regulators of Schizophyllum commune.Scientific reports · 2021Article
- Genetic Manipulation of the Brassicaceae Smut FungusJournal of fungi (Basel, Switzerland) · 2021Article
- Exploring molecular tools for transformation and gene expression in the cultivated edible mushroom Agrocybe aegerita.Molecular genetics and genomics : MGG · 2019Article
- High-throughput targeted gene deletion in the model mushroom Schizophyllum commune using pre-assembled Cas9 ribonucleoproteins.Scientific reports · 2019Article
- Nourseothricin N-acetyl transferase (NAT), a new selectable marker for nuclear gene expression inPlant methods · 2019Article
- Development of an Expression Vector to Overexpress or Downregulate Genes inJournal of fungi (Basel, Switzerland) · 2018Article
- Hydrophobin gene deletion and environmental growth conditions impact mechanical properties of mycelium by affecting the density of the material.Scientific reports · 2018Article
- Nuclear arms races: Experimental evolution for mating success in the mushroom-forming fungus Schizophyllum commune.PloS one · 2018Article
- Transcription factors of Schizophyllum commune involved in mushroom formation and modulation of vegetative growth.Scientific reports · 2017Article
- Genetic Manipulation of the Plant Pathogen Ustilago maydis to Study Fungal Biology and Plant Microbe Interactions.Journal of visualized experiments : JoVE · 2016Article
- Schizophyllum commune has an extensive and functional alternative splicing repertoire.Scientific reports · 2016Article
- Development in Aspergillus.Studies in mycology · 2013Article
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Phleomycin is mutagenic by introducing double-strand breaks in DNA. The ble gene of Streptoalloteychus hindustanus, which confers resistance to this substance, is widely used as a selection marker for transformation. Schizophyllum commune grows on 25 microg of phleomycin ml(-1) after introduction of a resistance cassette based on the ble gene. However, we here report that growth of resistant colonies on this concentration of phleomycin resulted in aberrant colony morphologies. Apparently, phleomycin was mutagenic despite acquired resistance. Therefore, a new selection system was developed based on resistance to the antibiotic nourseothricin. However, the transformation efficiency was tenfold lower than that obtained with phleomycin as a selection agent. This low transformation efficiency could be rescued by addition of a nonselective concentration of phleomycin during protoplast regeneration. This was accompanied by a higher incidence of single-copy integrations and with an increase of expression of key genes involved in double-strand break repair. Taken together, we conclude that the effect of a nonselective concentration of phleomycin strongly resembles the effect of restriction enzyme-mediated integration (REMI) but, unlike REMI, it does not depend on the presence of a target restriction site.
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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.