PMID:16002655

Citation	Krappmann, S, Bayram, O and Braus, GH (2005) Deletion and allelic exchange of the Aspergillus fumigatus veA locus via a novel recyclable marker module. Eukaryotic Cell 4:1298-307
Abstract	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 nitrate-containing medium, a phenotype that is completely restored in the reconstituted strain.
Links	PubMed PMC1168958 Online version:10.1128/EC.4.7.1298-1307.2005
Keywords	Alleles; Amino Acid Sequence; Aspergillus fumigatus/genetics; Conserved Sequence; Gene Deletion; Gene Expression Regulation, Fungal; Gene Targeting/methods; Genetic Markers/genetics; Molecular Sequence Data
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
ASPFU:VEA	acts_upstream_of_or_within	GO:0043936: asexual sporulation resulting in formation of a cellular spore	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
ASPFU:VEA	acts_upstream_of_or_within	GO:0043945: positive regulation of asexual sporulation resulting in formation of a cellular spore	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
ASPFM:Q6MYN4	involved_in	GO:0043945: positive regulation of asexual sporulation resulting in formation of a cellular spore	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete
ASPFM:Q6MYN4	involved_in	GO:0043936: asexual sporulation resulting in formation of a cellular spore	ECO:0000315: mutant phenotype evidence used in manual assertion		P		Seeded From UniProt	complete

References

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