ArticleMolecular genetics and genomics : MGG2004
Efficient gene disruption in the koji-mold Aspergillus sojae using a novel variation of the positive-negative method.
Article in Molecular genetics and genomics : MGG, 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 18 papers.
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18 citing papers in PubMed, 49 citations in OpenAlex.
- Article
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- Co-transformation of Aspergillus fumigatus: a simple and efficient strategy for gene editing without linking selectable markers.Access microbiology · 2025Article
- Phosphatidylcholine levels regulate hyphal elongation and differentiation in the filamentous fungus Aspergillus oryzae.Scientific reports · 2024Article
- Metabolic Engineering Techniques to Increase the Productivity of Primary and Secondary Metabolites Within Filamentous Fungi.Frontiers in fungal biology · 2021Article
- Disruption or reduced expression of the orotidine-5'-decarboxylase gene pyrG increases citric acid production: a new discovery during recyclable genome editing in Aspergillus niger.Microbial cell factories · 2020Article
- Translocated duplication of a targeted chromosomal segment enhances gene expression at the duplicated site and results in phenotypic changes inFungal biology and biotechnology · 2018Article
- Identification of a novel sesquiterpene biosynthetic machinery involved in astellolide biosynthesis.Scientific reports · 2016Article
- Targeted tandem duplication of a large chromosomal segment in Aspergillus oryzae.Applied and environmental microbiology · 2014Article
- Gene cloning, purification, and characterization of a novel peptidoglutaminase-asparaginase from Aspergillus sojae.Applied and environmental microbiology · 2012Article
- Analysis of the functions of recombination-related genes in the generation of large chromosomal deletions by loop-out recombination in Aspergillus oryzae.Eukaryotic cell · 2012Article
- A further study on chromosome minimization by protoplast fusion in Aspergillus oryzae.Molecular genetics and genomics : MGG · 2012Article
- SclR, a basic helix-loop-helix transcription factor, regulates hyphal morphology and promotes sclerotial formation in Aspergillus oryzae.Eukaryotic cell · 2011Article
- A trial of minimization of chromosome 7 in Aspergillus oryzae by multiple chromosomal deletions.Molecular genetics and genomics : MGG · 2010Article
- Identification of a basic helix-loop-helix-type transcription regulator gene in Aspergillus oryzae by systematically deleting large chromosomal segments.Applied and environmental microbiology · 2009Article
- Generation of large chromosomal deletions in koji molds Aspergillus oryzae and Aspergillus sojae via a loop-out recombination.Applied and environmental microbiology · 2008Article
- MpkA-Dependent and -independent cell wall integrity signaling in Aspergillus nidulans.Eukaryotic cell · 2007Article
- Enhanced gene targeting frequency in ku70 and ku80 disruption mutants of Aspergillus sojae and Aspergillus oryzae.Molecular genetics and genomics : MGG · 2006Article
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
When no phenotypic screen is available, gene disruption in the koji-mold Aspergillus sojae is a time-consuming process, owing to the low frequency of homologous recombination. To achieve efficient gene disruption in the koji-mold, we developed a novel positive-negative selection method to enrich for homologous recombinants. The pyrG gene from A. sojae was used as a positive selection marker for transformants, and the oliC31 gene of A. nidulans, which codes for a mutant form of subunit 9 of the F1FO-ATPase, was employed as a negative selection marker to facilitate elimination of non-homologous recombinants among the transformants. The positive-negative selection markers, in combination with a pyrG deletion strain as a host, enabled enrichment for homologous recombinants, and disruption of the genes niaD, areA and tannase was successfully demonstrated. In order to examine whether the positive-negative selection technique is effective for targeting any locus, even in the absence of information on gene function or phenotype, we attempted to disrupt the aflR gene of A. sojae, which codes for a putative transcription factor for the aflatoxin biosynthetic pathway, using the method. Despite the fact that this gene is not transcribed in A. sojae, aflR disruptants were efficiently obtained, suggesting that the method is indeed capable of targeting any locus, without additional ectopic integration, and is thus applicable for functional genomics studies in filamentous fungi, including A. sojae.
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