dc.contributor.author
Erdmann, Eileen A.
dc.contributor.author
Brandhorst, Antonia K. M.
dc.contributor.author
Gorbushina, Anna A.
dc.contributor.author
Schumacher, Julia
dc.date.accessioned
2024-09-05T07:54:57Z
dc.date.available
2024-09-05T07:54:57Z
dc.identifier.uri
https://refubium.fu-berlin.de/handle/fub188/44799
dc.identifier.uri
http://dx.doi.org/10.17169/refubium-44509
dc.description.abstract
Knufia petricola is a black fungus that colonizes sun-exposed surfaces as extreme and oligotrophic environments. As ecologically important heterotrophs and biofilm-formers on human-made surfaces, black fungi form one of the most resistant groups of biodeteriorating organisms. Due to its moderate growth rate in axenic culture and available protocols for its transformation and CRISPR/Cas9-mediated genome editing, K. petricola is used for studying the morpho-physiological adaptations shared by extremophilic and extremotolerant black fungi. In this study, the bacteria-derived tetracycline (TET)-dependent promoter (Tet-on) system was implemented to enable controllable gene expression in K. petricola. The functionality i.e., the dose-dependent inducibility of TET-regulated constructs was investigated by using GFP fluorescence, pigment synthesis (melanin and carotenoids) and restored uracil prototrophy as reporters. The newly generated cloning vectors containing the Tet-on construct, and the validated sites in the K. petricola genome for color-selectable or neutral insertion of expression constructs complete the reverse genetics toolbox. One or multiple genes can be expressed on demand from different genomic loci or from a single construct by using 2A self-cleaving peptides, e.g., for localizing proteins and protein complexes in the K. petricola cell or for using K. petricola as host for the expression of heterologous genes.
en
dc.format.extent
13 Seiten
dc.rights.uri
https://creativecommons.org/licenses/by/4.0/
dc.subject
Microcolonial fungi
en
dc.subject
Inducible promoter
en
dc.subject
Bimolecular fluorescence complementation
en
dc.subject
CRISPR/Cas9-mediated genome editing
en
dc.subject
Pigment synthesis
en
dc.subject.ddc
500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie::570 Biowissenschaften; Biologie
dc.title
The Tet-on system for controllable gene expression in the rock-inhabiting black fungus Knufia petricola
dc.type
Wissenschaftlicher Artikel
dcterms.bibliographicCitation.articlenumber
38
dcterms.bibliographicCitation.doi
10.1007/s00792-024-01354-2
dcterms.bibliographicCitation.journaltitle
Extremophiles
dcterms.bibliographicCitation.number
3
dcterms.bibliographicCitation.volume
28
dcterms.bibliographicCitation.url
https://doi.org/10.1007/s00792-024-01354-2
refubium.affiliation
Geowissenschaften
refubium.affiliation.other
Institut für Geologische Wissenschaften / Fachrichtung Geochemie, Hydrogeologie, Mineralogie
refubium.resourceType.isindependentpub
no
dcterms.accessRights.openaire
open access
dcterms.isPartOf.eissn
1433-4909
refubium.resourceType.provider
WoS-Alert