Construction of a codon-adapted Nourseotricin-resistance marker gene for efficient targeted gene deletion in the mycophenolic acid producer \(\textit {Penicillium brevicompactum}\)

  • \(\textit {Penicillium brevicompactum}\) is a filamentous ascomycete used in the pharmaceutical industry to produce mycophenolic acid, an immunosuppressant agent. To extend options for genetic engineering of this fungus, we have tested two resistance markers that have not previously been applied to \(\textit {P. brevicompactum}\). Although a generally available phleomycin resistance marker \(\it (ble)\) was successfully used in DNA-mediated transformation experiments, we were not able to use a commonly applicable nourseothricin resistance cassette \(\textit {(nat1)}\). To circumvent this failure, we constructed a new nat gene, considering the codon bias for \(\textit {P. brevicompactum}\). We then used this modified nat gene in subsequent transformation experiments for the targeted disruption of two nuclear genes, \(\textit {MAT1-2-1}\) and \(\it flbA\). For \(\textit {MAT1-2-1}\), we obtained deletion strains with a frequency of about 10%. In the case of flbA, the frequency was about 4%, and this disruption strain also showed reduced conidiospore formation. To confirm the deletion, we used \(\it ble\) to reintroduce the wild-type genes. This step restored the wild-type phenotype in the \(\it flbA\) deletion strain, which had a sporulation defect. The successful transformation system described here substantially extends options for genetically manipulating the biotechnologically relevant fungus \(\textit {P. brevicompactum}\).

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Metadaten
Author:Yasaman MahmoudjanlouGND, Birgit HoffGND, Ulrich KückORCiDGND
URN:urn:nbn:de:hbz:294-69418
DOI:https://doi.org/10.3390/jof5040096
Parent Title (English):Journal of Fungi
Publisher:MDPI
Place of publication:Basel
Document Type:Article
Language:English
Date of Publication (online):2020/02/05
Date of first Publication:2019/10/10
Publishing Institution:Ruhr-Universität Bochum, Universitätsbibliothek
Tag:MAT1-2-1; Penicillium brevicompactum; codon-adapted nourseothricin resistance gene; flbA; homologous recombination
Volume:5
Issue:4
First Page:96-1
Last Page:96-11
Institutes/Facilities:Lehrstuhl für Allgemeine und Molekulare Botanik
Dewey Decimal Classification:Naturwissenschaften und Mathematik / Pflanzen (Botanik)
open_access (DINI-Set):open_access
faculties:Fakultät für Biologie und Biotechnologie
Licence (English):License LogoCreative Commons - CC BY 4.0 - Attribution 4.0 International