Abstract
We disrupted the Aspergillus fumigatus argB gene, encoding ornithine transcarbamylase, using a novel in vitro transposon-based mutagenesis approach. This approach utilizes a modified transposon containing the Neurospora crassa pyr4 gene, which is randomly inserted in vitro into a target sequence of interest. Clones in which the gene of interest has been disrupted are identified by PCR and used to transform a pyrG-deficient strain of A. fumigatus. Using this approach, we obtained arginine auxotrophs of A. fumigatus. Full characterization of the argB insertion was performed by Southern blot analysis. These strains can be supplemented by addition of arginine into the culture medium and can be fully rescued to arginine prototrophy by transformation with the intact A. fumigatus argB gene.
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Acknowledgements
We thank Dr. Doron Goldberg for his help in the preparation of the alignments in Fig. 1. This work was funded by the Israel Academy of Sciences grant 741/01 to N.O.
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Communicated by U. Kück
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Jadoun, J., Shadkchan, Y. & Osherov, N. Disruption of the Aspergillus fumigatus argB gene using a novel in vitro transposon-based mutagenesis approach. Curr Genet 45, 235–241 (2004). https://doi.org/10.1007/s00294-003-0480-6
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DOI: https://doi.org/10.1007/s00294-003-0480-6