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PMID:16002655

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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

Significance

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

Notes

See also

References

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