Deletion and Allelic Exchange of the Aspergillus fumigatus veA Locus via a Novel Recyclable Marker Module

Krappman, Sven and Bayram, Ozgur and Braus, Gerhard H. (2005) Deletion and Allelic Exchange of the Aspergillus fumigatus veA Locus via a Novel Recyclable Marker Module. Eukaryotic Cell, 4 (7). pp. 1298-1307. ISSN 1535-9778

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Detailed evaluation of gene functions in an asexual fungus requires advanced methods of molecular biology. For the generation of targeted gene deletions in the opportunistic pathogen Aspergillus fumigatus we designed a novel blaster module allowing dominant selection of transformants due to resistance to phleomycin as well as dominant (counter)selection of a Cre recombinase-mediated marker excision event. For validation purposes we have deleted the A. fumigatus pabaA gene in a wild-type isolate by making use of this cassette. The resulting pabaA::loxP strain served as the recipient for subsequent targeting of the velvet locus. Homologous reconstitution of the deleted gene was performed by an allele whose expression is driven in a nitrogen source-dependent manner, as validated by Northern analyses. Overexpression of the veA locus in A. fumigatus does not result in any obvious phenotype, whereas the sporulation capacities of the veA null mutant are reduced on nitratecontaining medium, a phenotype that is completely restored in the reconstituted strain.

Item Type: Article
Keywords: Deletion; Allelic Exchange; Aspergillus fumigatus; veA Locus; Novel Recyclable Marker Module;
Academic Unit: Faculty of Science and Engineering > Biology
Item ID: 6249
Identification Number: 10.1128/EC.4.7.1298–1307.2005
Depositing User: Ozgur Bayram
Date Deposited: 14 Jul 2015 15:32
Journal or Publication Title: Eukaryotic Cell
Publisher: American Society for Microbiology
Refereed: Yes

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