Selection of multiple disruption events in Aspergillus fumigatus using the orotidine-5′-decarboxylase gene, pyrG, as a unique transformation marker

d'Enfert, Christophe

doi:10.1007/s002940050103

Cited by 178Fungal and yeast genetics researchAntifungal resistance and susceptibilityBacterial Genetics and Biotechnology

Selection of multiple disruption events in Aspergillus fumigatus using the orotidine-5′-decarboxylase gene, pyrG, as a unique transformation marker

d'Enfert, Christophe·1996·Current Genetics

DOI 10.1007/s002940050103

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Abstract

A 8.6-kb disruption cassette, referred to here as a pyrG-blaster and consisting of the Aspergillus niger pyrG gene flanked by a direct repeat that encodes the neomycin phosphotransferase of transposon Tn5 was constructed. Following transformation of a uridine/uracil auxotrophic pyrG strain of A. fumigatus, genomic insertions of the pyrG-blaster were obtained either by targeted gene replacement at the rodA locus, resulting in the formation of hydrophilic spores, or by ectopic integration. In both cases, recombination between the two elements of the direct repeat could be selected in the presence of 5-fluoro-orotic acid and resulted in the excision of the A. niger pyrG gene, producing A. fumigatus uridine/uracil auxotrophs that retained their additional mutant phenotype because of the persistence of one of the two elements of the direct repeat at the site of insertion of the pyrG-blaster. Selection for uracil/uridine prototrophy can therefore be used again to disrupt another gene.

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A. fumigatus strain CEA17 is a uracil-auxotrophic (pyrG1) mutant of A. fumigatus strain CEA10.
“A. fumigatus strain CEA17 is a uracil-auxotrophic (pyrG1) mutant of A. fumigatus strain CEA10 [67],[68].”
From A Sterol-Regulatory Element Binding Protein Is Required for Cell Polarity, Hypoxia Adaptation, Azole Drug Resistance, and Virulence in Aspergillus fumigatus (2008) · cites this paper as [68]

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Authors: d'Enfert, C.
Venue: Current Genetics
Publisher: Springer Science and Business Media LLC
Published: 1996-06
Volume: 30
Issue: 1
Pages: 76-82
ISSN: 0172-8083 (ISSN-L)
1432-0983
DOI: 10.1007/s002940050103
OpenAlex: W2165326582
Cited by: 178
References: 33
Citations by year: 2025: 4 · 2024: 5 · 2023: 5 · 2022: 3 · 2021: 13 · 2020: 10 · 2019: 11 · 2018: 7 · 2017: 4 · 2016: 1 · 2015: 10 · 2014: 4 · 2013: 3 · 2012: 8
FWCI: 1.01
Type: journal article
Language: en
Open access: closed
Licence: www.springernature.com/gp/researchers/text-and-data-mining (tdm)
www.springernature.com/gp/researchers/text-and-data-mining (vor)
Locations: Current Genetics (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Health Sciences › Medicine › Infectious Diseases › Antifungal resistance and susceptibility
Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
Keywords: Aspergillus fumigatus, pyrG, gene disruption, Tn5 transposon, neomycin phosphotransferase, ectopic integration, targeted gene replacement
MeSH terms: Aspergillus fumigatus, Base Sequence, DNA, Fungal, Genes, Fungal, Genetic Markers, Molecular Sequence Data, Orotidine-5'-Phosphate Decarboxylase, Transformation, Genetic, Restriction Mapping, Mutagenesis, Insertional

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Selection of multiple disruption events in Aspergillus fumigatus using the orotidine-5′-decarboxylase gene, pyrG, as a unique transformation marker

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