Genetic transformation system for the aflatoxin-producing fungus Aspergillus flavus
| Autor: | E R Seip, C P Woloshuk, Gary A. Payne, C. R. Adkins |
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| Rok vydání: | 1989 |
| Předmět: |
Genetic Vectors
Aspergillus flavus Biology Applied Microbiology and Biotechnology Parasexual cycle Microbiology Neurospora crassa chemistry.chemical_compound Plasmid Transformation Genetic Aflatoxins heterocyclic compounds DNA Fungal Uracil Ecology Genetic transfer fungi food and beverages Nucleic Acid Hybridization biology.organism_classification Cosmids Culture Media Transformation (genetics) chemistry Mutation Cosmid DNA Food Science Biotechnology Plasmids Research Article |
| Zdroj: | Applied and environmental microbiology. 55(1) |
| ISSN: | 0099-2240 |
| Popis: | A heterologous transformation system was developed for Aspergillus flavus with efficiencies greater than 20 stable transformants per micrograms of DNA. Protoplasts of uracil-requiring strains of the fungus were transformed with plasmid and cosmid vectors containing the pyr-4 gene of Neurospora crassa. Transformants were selected for their ability to grow and sporulate on medium lacking uracil. Vector DNA appeared to integrate randomly into the genome of A. flavus with a tendency for multiple, tandem insertion. Transformants with single or multiple insertions were stable after five consecutive transfers on medium containing uracil. Uracil-requiring recipient strains were obtained either by UV-irradiating conidia and selecting colonies resistant to 5-fluoroorotic acid or by transferring the mutated pyr locus to strains by parasexual recombination. This is the first report of a transformation system for an aflatoxin-producing fungus. The transformation system and the availability of aflatoxin-negative mutants provide a new approach to studying the biosynthesis and regulation of aflatoxin. |
| Databáze: | OpenAIRE |
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