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PMID: 2495764 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Genetic transformation system for the aflatoxin-producing fungus Aspergillus flavus.

Applied and environmental microbiology ·Vol. 55 ·No. 1 ·1989-01-00 ·Pages 86-90

Woloshuk CP, Seip ER, Payne GA, Adkins CR

Abstract

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.

MeSH Terms
Aflatoxins/biosynthesis Aspergillus flavus/genetics,growth & development,metabolism Cosmids Culture Media DNA, Fungal/genetics Genetic Vectors Mutation Nucleic Acid Hybridization Plasmids Transformation, Genetic Uracil/metabolism
Chemicals
Aflatoxins Culture Media DNA, Fungal Uracil
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Woloshuk C P
Department of Plant Pathology, North Carolina State University, Raleigh 27695-7616.
Seip E R
Payne G A
Adkins C R
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Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.55.1.86-90.1989
Published
1989-01-00
Pages
86-90
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC184058
Subset
IM
Grants
NCRR NIH HHS · RR 7071 · United States
External Links
PubMed source DOI: 10.1128/aem.55.1.86-90.1989 PMC Full Text View journal details
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