The Aspergillus nidulans amdS gene as a marker for the identification of multicopy T-DNA integration events in Agrobacterium-mediated transformation of Aspergillus awamori

article


Michielse, C.B.

Ram, A.F.J.

Hondel, C.A.M.J.J. van den
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.
Topics

















































BiotechnologyAgrobacterium tumefaciensAmdS selection markerAspergillus awamoriFilamentous fungiMulticopy T-DNA integrationTransformationAcetamidaseAcetamideAgarDNAFungal enzymeHygromycinNitrogenUnclassified drugAgar mediumAnalytic methodAntibiotic resistanceAspergillus awamoriAspergillus nidulansControlled studyCulture mediumDNA integrationFrequency analysisFungus identificationGene transferGenetic selectionGenetic transformationGrowth rateIntermethod comparisonMarker geneNonhumanPriority journalRhizobiumAmidohydrolasesAspergillusCinnamatesGene Expression Regulation, FungalGenes, FungalGenetic MarkersHygromycin BPlasmidsRhizobiumTransformation, GeneticAgrobacteriumAgrobacterium tumefaciensAspergillusAspergillus awamoriEmericella nidulansFungi
TNO Identifier
237834
Repository link
https://resolver.tno.nl/uuid:ef50b64c-246a-467e-bb69-b5e4f09cc2c0
ISSN
01728083
Source
Current Genetics, 45(6), pp. 399-403.
Pages
399-403
Files
To receive the publication files, please send an e-mail request to TNO Repository.