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Article

Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation

Institute of Molecular Biology, Biocenter, Medical University of Innsbruck, 6020 Innsbruck, Austria
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Academic Editor: Jürgen Löffler
J. Fungi 2021, 7(7), 506; https://doi.org/10.3390/jof7070506
Received: 19 May 2021 / Revised: 17 June 2021 / Accepted: 18 June 2021 / Published: 24 June 2021
(This article belongs to the Section Fungal Genomics, Genetics and Molecular Biology)
The hygromycin B phosphotransferase gene from Escherichia coli and the pyrithiamine resistance gene from Aspergillus oryzae are two dominant selectable marker genes widely used to genetically manipulate several fungal species. Despite the recent development of CRISPR/Cas9 and marker-free systems, in vitro molecular tools to study Aspergillus fumigatus, which is a saprophytic fungus causing life-threatening diseases in immunocompromised hosts, still rely extensively on the use of dominant selectable markers. The limited number of drug selectable markers is already a critical aspect, but the possibility that their introduction into a microorganism could induce enhanced virulence or undesired effects on metabolic behavior constitutes another problem. In this context, here, we demonstrate that the use of ptrA in A. fumigatus leads to the secretion of a compound that allows the recovery of thiamine auxotrophy. In this study, we developed a simple modification of the two commonly used dominant markers in which the development of resistance can be controlled by the xylose-inducible promoter PxylP from Penicillium chrysogenum. This strategy provides an easy solution to avoid undesired side effects, since the marker expression can be readily silenced when not required. View Full-Text
Keywords: dominant selectable marker; Aspergillus fumigatus; inducible marker; hph; ptrA; xylanase promoter; thiamine; hygromycin; pyrithiamine dominant selectable marker; Aspergillus fumigatus; inducible marker; hph; ptrA; xylanase promoter; thiamine; hygromycin; pyrithiamine
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MDPI and ACS Style

Baldin, C.; Kühbacher, A.; Merschak, P.; Sastré-Velásquez, L.E.; Abt, B.; Dietl, A.-M.; Haas, H.; Gsaller, F. Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation. J. Fungi 2021, 7, 506. https://doi.org/10.3390/jof7070506

AMA Style

Baldin C, Kühbacher A, Merschak P, Sastré-Velásquez LE, Abt B, Dietl A-M, Haas H, Gsaller F. Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation. Journal of Fungi. 2021; 7(7):506. https://doi.org/10.3390/jof7070506

Chicago/Turabian Style

Baldin, Clara, Alexander Kühbacher, Petra Merschak, Luis E. Sastré-Velásquez, Beate Abt, Anna-Maria Dietl, Hubertus Haas, and Fabio Gsaller. 2021. "Inducible Selectable Marker Genes to Improve Aspergillus fumigatus Genetic Manipulation" Journal of Fungi 7, no. 7: 506. https://doi.org/10.3390/jof7070506

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