Genetic Manipulation of Aspergillus Nidulans: Heterokaryons and Diploids for Dominance, Complementation and Haploidization Analyses

Nat Protoc. 2007;2(4):822-30. doi: 10.1038/nprot.2007.113.

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.

MeSH terms

  • Aspergillus nidulans / classification
  • Aspergillus nidulans / genetics*
  • Aspergillus nidulans / physiology
  • Chromosome Mapping
  • Diploidy
  • Genes, Dominant*
  • Genes, Fungal
  • Genetic Complementation Test / methods*
  • Genetic Engineering / methods*
  • Genotype
  • Haploidy*
  • Heterozygote
  • Mutation