Open AccessExpression of the red fluorescent protein DsRed‐Express in filamentous ascomycete fungi
Author(s) -
Mikkelsen Lisbeth,
Sarrocco Sabrina,
Lübeck Mette,
Jensen Dan Funck
Publication year - 2003
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1016/s0378-1097(03)00355-0
Subject(s) - aspergillus nidulans , biology , green fluorescent protein , expression vector , transformation (genetics) , hygromycin b , gene , microbiology and biotechnology , recombinant dna , genetics , mutant
The recently reported red fluorescent protein DsRed from the reef coral Discosoma sp. represents a new marker that has been codon‐optimized for high expression in mammalian cells. To facilitate expression of DsRed in ascomycete fungi, we used the clone pDsRed‐Express (Clontech) for constructing a plasmid vector, pPgpd‐DsRed, containing the constitutive Aspergillus nidulans glyceraldehyde 3‐phosphate ( gpd ) promoter. This vector was used for co‐transformation of Penicillium paxilli , Trichoderma harzianum and Trichoderma virens (syn. Gliocladium virens ) together with either pAN7‐1 or gGFP, both containing a gene for hygromycin resistance for transformant selection. In addition, gGFP contains a green fluorescent protein (GFP) gene for expression in Ascomycetes. Expression of DsRed‐Express was obtained in all three fungi, indicating that DsRed can be used as a highly effective vital marker in Ascomycetes. Dual marked transformants expressed both DsRed‐Express and GFP in the same mycelium and were used for non‐quantitative comparison of the intensity of the fluorescence using confocal laser scanning microscopy.