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Genetic manipulation of Aspergillus nidulans: heterokaryons and diploids for dominance, complementation and haploidization analyses

Nature Protocols volume 2, pages 822–830 (2007)Cite this article

Abstract

The haploid microbial eukaryote Aspergillus nidulans is a powerful genetic system, which allows analysis of a broad range of biological phenomena. In addition to conventional analysis of meiotic progeny in a single generation, parasexual analysis affords a rapid and convenient method for genetic analysis. We describe the construction of A. nidulans heterokaryons and diploids for use in genetic analysis to determine dominance and conduct complementation tests. We also describe the rapid mapping of mutations to chromosomes by haploidization of diploids carrying marked chromosomes. Balanced heterokaryons may be established within 10 days and diploids may be constructed in 2–3 weeks. Dominance tests and complementation tests using balanced heterokaryons or diploids may be completed in 2–3 days. Haploidization analysis of heterozygous diploids can be achieved within 10 days. These protocols should be adaptable for use in related Aspergilli and Penicillia, which lack a known meiotic cycle.

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Figure 1: Life cycle of A. nidulans. A. nidulans grows as haploid vegetative filamentous hyphae following germination of uninucleate conidia or binucleate ascospores.
Figure 2: Tests for dominance and complementation.
Figure 3: Segregation of markers during haploidization of A. nidulans heterozygous diploids.
Figure 4: Assignment of a mutation to a chromosome by haploidization of an A. nidulans heterozygous diploid.
Figure 5: Assignment of a reciprocal translocation to chromosomes by haploidization of an A. nidulans heterozygous diploid.
Figure 6: Construction of A. nidulans heterokaryons and diploids.
Figure 7: Haploidization of A. nidulans diploids.

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Acknowledgements

We thank Q. Lang for photography and K. Nguygen for technical assistance.

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  1. Department of Genetics, The University of Melbourne, Parkville, 3010, Victoria, Australia

    Richard B Todd, Meryl A Davis & Michael J Hynes

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  1. Richard B Todd
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  3. Michael J Hynes
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Correspondence to Richard B Todd.

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Todd, R., Davis, M. & Hynes, M. Genetic manipulation of Aspergillus nidulans: heterokaryons and diploids for dominance, complementation and haploidization analyses. Nat Protoc 2, 822–830 (2007). https://doi.org/10.1038/nprot.2007.113

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