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Published on: February 15, 2019
Isogenic auxotrophic mutant strains in the Aspergillus fumigatus genome reference strain AF293
Tao Xue1, Cuong K Nguyen, Angela Romans
1The Genes and Development Graduate Program, Division of Pathology and Laboratory Medicine, The University of Texas Graduate School of Biomedical Sciences, The University of Texas M. D. Anderson Cancer Center, 1515 Holcombe Blvd., Houston, TX 77030, USA.
Abstract:
Aspergillus fumigatus is a ubiquitous fungus that is a frequent opportunistic pathogen in immunosuppressed patients. Because of its role as a pathogen, it is of considerable experimental interest. A set of auxotrophic isogenic strains in the A. fumigatus genome reference strain AF293 has been developed. Using molecular genetic methods, arginine and lysine auxotrophs were made by deletion of argB and lysB, respectively. Transformation of these auxotrophic strains with plasmids carrying argB or lysB, respectively, results in efficient integration at these loci. Finally, these strains are able to form stable diploids, which should further facilitate analysis of gene functions in this fungus. Furthermore, the development of this isogenic set of auxotrophic strains in the AF293 background will enable investigators to study this important opportunistic human pathogen with greater facility.
Insights
Researchers developed new Aspergillus fumigatus strains for studying this opportunistic pathogen. These isogenic auxotrophic strains facilitate genetic analysis and diploid formation in A. fumigatus research.
Area of Science:
- Medical Mycology
- Molecular Genetics
- Fungal Pathogenesis
Background:
- Aspergillus fumigatus is a common opportunistic pathogen, particularly in immunocompromised individuals.
- Its significant role in human health necessitates advanced research tools for understanding its pathogenicity.
Purpose of the Study:
- To develop a set of isogenic auxotrophic strains of Aspergillus fumigatus in the AF293 reference background.
- To facilitate experimental analysis of gene functions and pathogenicity mechanisms in A. fumigatus.
Main Methods:
- Deletion of the argB and lysB genes to create arginine and lysine auxotrophs, respectively.
- Transformation of auxotrophic strains with plasmids for gene reintroduction and locus integration.
- Induction of stable diploid formation in the engineered strains.
Main Results:
- Successfully generated arginine and lysine auxotrophic isogenic strains of A. fumigatus.
- Demonstrated efficient integration of reintroduced genes at the target loci.
- Established stable diploid strains, enhancing genetic analysis capabilities.
Conclusions:
- The developed isogenic auxotrophic strains provide a valuable resource for studying Aspergillus fumigatus.
- These tools will significantly improve the ease and depth of genetic studies on this opportunistic human pathogen.
