Title
The Aspergillus nidulans amdS gene as a marker for the identification of multicopy T-DNA integration events in Agrobacterium-mediated transformation of Aspergillus awamori
Author
Michielse, C.B.
Ram, A.F.J.
van den Hondel, C.A.M.J.J.
TNO Voeding
Publication year
2004
Abstract
The Aspergillus nidulans amdS selection marker was used for the identification of multicopy T-DNA insertions in Agrobacterium-mediated transformation of Asp. awamori. The selection of transformants on agar plates containing acetamide as sole nitrogen source and hygromycin resulted in a six-fold decrease in the transformation frequency, compared with the transformation frequency obtained after hygromycin selection alone. However, it was found that 47% of the transformants obtained after hygromycin and acetamide double selection contained multiple T-DNA integrations. Furthermore, it was found that the multicopy transformants could easily be identified based on their growth rate on agar plates containing acetamide medium. Based on these data, it can be concluded that the amdS marker can also be used as a selection marker in Agrobacterium-mediated transformation of Asp. awamori and that it is a very useful marker to identify those transformants containing multiple T-DNA integrations. © Springer-Verlag 2004.
Subject
Biology
Biotechnology
Agrobacterium tumefaciens
amdS selection marker
Aspergillus awamori
Filamentous fungi
Multicopy T-DNA integration
Transformation
acetamidase
acetamide
agar
DNA
fungal enzyme
hygromycin
nitrogen
unclassified drug
agar medium
analytic method
antibiotic resistance
article
Aspergillus awamori
Aspergillus nidulans
controlled study
culture medium
DNA integration
frequency analysis
fungus identification
gene transfer
genetic selection
genetic transformation
growth rate
intermethod comparison
marker gene
nonhuman
priority journal
Rhizobium
Amidohydrolases
Aspergillus
Cinnamates
Gene Expression Regulation, Fungal
Genes, Fungal
Genetic Markers
Hygromycin B
Plasmids
Rhizobium
Transformation, Genetic
Agrobacterium
Agrobacterium tumefaciens
Aspergillus
Aspergillus awamori
Emericella nidulans
Fungi
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DOI
https://doi.org/10.1007/s00294-004-0500-1
TNO identifier
237834
ISSN
0172-8083
Source
Current Genetics, 45 (6), 399-403
Document type
article