Journal
FUNGAL GENETICS AND BIOLOGY
Volume 46, Issue 1, Pages 67-76Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.fgb.2008.10.005
Keywords
Carboxin; Aspergillus oryzae; Succinate dehydrogenase; Transformation; Dominant selectable marker
Categories
Funding
- Ministry of Education, Science and Culture of Japan
- NARO, Japan
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Mutants exhibiting resistance to the fungicide, carboxin, were isolated from Aspergillus oryzae, and the mutations in the three gene loci, which encode succinate dehydrogenase (SDH) B, C, and D subunits, were identified to be independently responsible for the resistance. A structural model of the SDH revealed the different mechanisms that confer carboxin-resistance in different mutations. The mutant AosdhB gene (AosdhB(cxr)) Was further examined for possible use as a transformant selection marker. After transformation with AosdhB(cxr), carboxin-resistant colonies appeared within 4 days of culture, and all of the examined colonies carried the transgene. Insertion analyses revealed that the AosdhB(cxr) gene was integrated into AosdhB locus via homologous recombination at high efficiency. Furthermore, AosdhB(cxr) functioned as a successful selection marker in a transformation experiment in Aspergillus parasiticus, Suggesting that this transformation system can be used for Aspergillus species. (C) 2008 Elsevier Inc. All rights reserved.
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