The construction and use of versatile binary vectors carrying pyrG auxotrophic marker and fluorescent reporter genes for Agrobacterium-mediated transformation of Aspergillus oryzae

Aspergillus oryzae is a safe mold widely used in food industry. It is also considered as a microbial cell factory for production of recombinant proteins and enzymes. Currently, genetic manipulation of filamentous fungi is achieved via Agrobacterium tumefaciens -mediated transformation methods usuall...

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Bibliographic Details
Published in:World journal of microbiology & biotechnology 2016-12, Vol.32 (12), p.204-204, Article 204
Main Authors: Nguyen, Khuyen Thi, Ho, Quynh Ngoc, Pham, Thu Ha, Phan, Tuan-Nghia, Tran, Van-Tuan
Format: Article
Language:English
Subjects:
Agrobacterium tumefaciens - physiology
Antibiotics
Applied Microbiology
Aspergillus oryzae - genetics
Biochemistry
Biomedical and Life Sciences
Biosynthesis
Biotechnology
Drug resistance
Environmental Engineering/Biotechnology
Enzymes
Fungi
Gene expression
Genes, Fungal
Genes, Reporter
Genetic engineering
Genetic Vectors - genetics
Genotype & phenotype
Glucose
Laboratories
Life Sciences
Microbiology
Original Paper
Plasmids
Polyethylene glycol
Promoter Regions, Genetic
Protein Engineering
Proteins
Recombinant Proteins - metabolism
Transformation, Genetic
Virulence
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Summary:Aspergillus oryzae is a safe mold widely used in food industry. It is also considered as a microbial cell factory for production of recombinant proteins and enzymes. Currently, genetic manipulation of filamentous fungi is achieved via Agrobacterium tumefaciens -mediated transformation methods usually employing antibiotic resistance markers. These methods are hardly usable for A. oryzae due to its strong resistance to the common antifungal compounds used for fungal transformation. In this study, we have constructed two binary vectors carrying the pyrG gene from A. oryzae as a biochemical marker than an antibiotic resistance marker, and an expression cassette for GFP or DsRed reporter gene under control of the constitutive gpdA promoter from Aspergillus nidulans . All components of these vectors are changeable to generate new versions for specific research purposes. The developed vectors are fully functional for heterologous expression of the GFP and DsRed fluorescent proteins in the uridine/uracil auxotrophic A. oryzae strain. Our study provides a new approach for A. oryzae transformation using pyrG as the selectable auxotrophic marker, A. tumefaciens as the DNA transfer tool and fungal spores as the transformation material. The binary vectors constructed can be used for gene expression studies in this industrially important filamentous fungus.
ISSN:0959-3993
1573-0972
DOI:10.1007/s11274-016-2168-3