ArticleCurrent genetics2009
Investigating dominant selection markers for Coprinopsis cinerea: a carboxin resistance system and re-evaluation of hygromycin and phleomycin resistance vectors.
Article in Current genetics, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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14 citing papers in PubMed, 24 citations in OpenAlex.
- Selection markers for transformation of the sequenced reference monokaryon Okayama 7/#130 and homokaryon AmutBmut ofFungal biology and biotechnology · 2020Article
- Exploring molecular tools for transformation and gene expression in the cultivated edible mushroom Agrocybe aegerita.Molecular genetics and genomics : MGG · 2019Article
- TrichoGate: An Improved Vector System for a Large Scale of Functional Analysis ofFrontiers in microbiology · 2019Article
- A silver bullet in a golden age of functional genomics: the impact ofFungal biology and biotechnology · 2017Review
- Development of a novel strategy for fungal transformation based on a mutant locus conferring carboxin-resistance in Magnaporthe oryzae.AMB Express · 2016Article
- The good, the bad and the tasty: The many roles of mushrooms.Studies in mycology · 2016Article
- Yeast recombination-based cloning as an efficient way of constructing vectors for Zymoseptoria tritici.Fungal genetics and biology : FG & B · 2015Article
- A gene locus for targeted ectopic gene integration in Zymoseptoria tritici.Fungal genetics and biology : FG & B · 2015Article
- Genetic engineering of Ganoderma lucidum for the efficient production of ganoderic acids.Bioengineered · 2015Article
- Introns regulate gene expression in Cryptococcus neoformans in a Pab2p dependent pathway.PLoS genetics · 2013Article
- Enhancement of ganoderic acid accumulation by overexpression of an N-terminally truncated 3-hydroxy-3-methylglutaryl coenzyme A reductase gene in the basidiomycete Ganoderma lucidum.Applied and environmental microbiology · 2012Article
- Mutagenesis and functional studies with succinate dehydrogenase inhibitors in the wheat pathogen Mycosphaerella graminicola.PloS one · 2012Article
- Investigating Agrobacterium-mediated transformation of Verticillium albo-atrum on plant surfaces.PloS one · 2010Article
- Establishing molecular tools for genetic manipulation of the pleuromutilin-producing fungus Clitopilus passeckerianus.Applied and environmental microbiology · 2009Article
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Authors and funding
6 authors at 3 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Dominant selectable markers are beneficial for transformation of many fungi, particularly those model species where repeated transformations may be required. A carboxin resistance allele of the Coprinopsis cinerea sdi1 gene, encoding the iron-sulphur protein subunit of succinate dehydrogenase, was developed by introducing a suitable point mutation in the histidine block responsible for binding of the associated iron ion. This modified gene was used successfully to confer carboxin resistance upon transformation of C. cinerea protoplasts. Plasmids previously used to establish hygromycin transformation systems of several basidiomycete species, such as pAN7-1 and phph004, failed to give rise to hygromycin-resistant transformants of C. cinerea, whilst pPHT1 was successful. Sequencing of these constructs showed that the hygromycin resistance gene in pAN7-1 and phph004 had been modified removing the codons encoding two lysine residues following the N-terminal methionine. Replacement of the deleted 6 bp (AAA AAG) in the truncated hph gene led to generation of hygromycin-resistant transformants indicating the importance of these two codons for expression in C. cinerea. Phleomycin-resistant (ble) transformants were also obtained, but only with the intron-containing construct pblei004, showing that an intron is necessary to obtain phleomycin-resistant C. cinerea. This contrasts with hygromycin-resistance, where introns are not required for expression, emphasising the variability in importance of these elements.
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