Development of a homologous transformation system for the human pathogenic fungus Aspergillus fumigatus based on the pyrG gene encoding orotidine 5'-monophosphate decarboxylase

Curr Genet. 1998 May;33(5):378-85. doi: 10.1007/s002940050350.


A homologous transformation system for the opportunistic fungal pathogen Aspergillus fumigatus was developed. It is based on the A. fumigatus pyrG gene, encoding orotidine 5'-monophosphate decarboxylase, which was cloned and sequenced. Transformation of both Aspergillus (Emericella) nidulans and A. fumigatus pyrG mutant strains by the use of protoplasts or electroporation established the functionality of the cloned gene. DNA sequencing of the A. fumigatus pyrG1 mutant allele revealed that it encodes a truncated, non-functional, PyrG protein. Transformation of an A. fumigatus pyrG1 mutant with a plasmid carrying the novel pyrG2 allele constructed by in vitro mutagenesis yielded prototrophic transformants following recombination between both mutation sites. Analysis of transformants carrying the entire plasmid showed that up to 45% of integration had occurred at the pyrG locus. This provides a tool to target defined genetic constructs at a specific locus in the A. fumigatus genome in order to study gene regulation and function.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aspergillus fumigatus / enzymology
  • Aspergillus fumigatus / genetics*
  • Aspergillus fumigatus / pathogenicity
  • Base Sequence
  • Cloning, Molecular
  • Consensus Sequence
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • Genes, Fungal*
  • Genetic Complementation Test
  • Humans
  • Molecular Sequence Data
  • Orotidine-5'-Phosphate Decarboxylase / biosynthesis
  • Orotidine-5'-Phosphate Decarboxylase / chemistry
  • Orotidine-5'-Phosphate Decarboxylase / genetics*
  • Polymerase Chain Reaction
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Sequence Alignment
  • Sequence Homology, Amino Acid
  • Transformation, Genetic*


  • DNA, Fungal
  • Recombinant Proteins
  • Orotidine-5'-Phosphate Decarboxylase