Characterization of the Aspergillus nidulans biotin biosynthetic gene cluster and use of the bioDA gene as a new transformation marker

Pasqualina Magliano, M. Flipphi, Dominique Sanglard, Yves Poirier

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The genes involved in the biosynthesis of biotin were identified in the hyphal fungus Aspergillus nidulans through homology searches and complementation of Escherichia coli biotin-auxotrophic mutants. Whereas the 7,8-diaminopelargonic acid synthase and dethiobiotin synthetase are encoded by distinct genes in bacteria and the yeast Saccharomyces cerevisiae, both activities are performed in A. nidulans by a single enzyme, encoded by the bifunctional gene bioDA. Such a bifunctional bioDA gene is a genetic feature common to numerous members of the ascomycete filamentous fungi and basidiomycetes, as well as in plants and oömycota. However, unlike in other eukaryota, the three bio genes contributing to the four enzymatic steps from pimeloyl-CoA to biotin are organized in a gene cluster in pezizomycotina. The A. nidulans auxotrophic mutants biA1, biA2 and biA3 were all found to have mutations in the 7,8-diaminopelargonic acid synthase domain of the bioDA gene. Although biotin auxotrophy is an inconvenient marker in classical genetic manipulations due to cross-feeding of biotin, transformation of the biA1 mutant with the bioDA gene from either A. nidulans or Aspergillus fumigatus led to the recovery of well-defined biotin-prototrophic colonies. The usefulness of bioDA gene as a novel and robust transformation marker was demonstrated in co-transformation experiments with a green fluorescent protein reporter, and in the efficient deletion of the laccase (yA) gene via homologous recombination in a mutant lacking non-homologous end-joining activity.

Original languageEnglish
Pages (from-to)208-215
Number of pages8
JournalFungal Genetics and Biology
Volume48
Issue number2
DOIs
Publication statusPublished - Feb 2011

Fingerprint

Aspergillus nidulans
Multigene Family
Biotin
Genes
dethiobiotin synthetase
Fungi
Laccase
Basidiomycota
Ascomycota
Aspergillus fumigatus
Homologous Recombination
Green Fluorescent Proteins
Eukaryota
Saccharomyces cerevisiae
Yeasts
Escherichia coli
Bacteria
Mutation

Keywords

  • Aspergillus nidulans
  • Biotin
  • Transformation

ASJC Scopus subject areas

  • Genetics
  • Microbiology

Cite this

Characterization of the Aspergillus nidulans biotin biosynthetic gene cluster and use of the bioDA gene as a new transformation marker. / Magliano, Pasqualina; Flipphi, M.; Sanglard, Dominique; Poirier, Yves.

In: Fungal Genetics and Biology, Vol. 48, No. 2, 02.2011, p. 208-215.

Research output: Contribution to journalArticle

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