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Generation and confirmation of PdcA and AlcC null mutants in A. fumigatus.

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posted on 21.07.2011, 01:20 authored by Nora Grahl, Srisombat Puttikamonkul, Jeffrey M. Macdonald, Michael P. Gamcsik, Lisa Y. Ngo, Tobias M. Hohl, Robert A. Cramer

Schematic of wild type (CEA10), PdcA (A), and AlcC (B) null mutant genomic loci. (A) Southern blot analysis of wild type, ΔpdcA, and pdcA recon strains as well as (B) wild type, ΔalcC, and alcC recon strains. Genomic DNA from the respective strains was isolated and digested overnight with SalI and NcoI in the case of PdcA and with only NcoI for AlcC. An approximate 1 kb genomic region was utilized as a probe. The expected hybridization patterns and sizes were observed for all strains tested. In addition, confirmation of ectopic reconstitution was confirmed by the presence of the wild type locus hybridization signal and persistence of the null mutant locus. The alcC reconstituted strain showed a double re-insertion of the alcC locus.

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