Expression of the red fluorescent protein DsRed-Express in filamentous ascomycete fungi.
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.
Mikkelsen
Lisbeth
L
Section of Plant Pathology, Department of Plant Biology, The Royal Veterinary and Agricultural University, 40 Thorvaldsensvej, DK-1871 C, Frederiksberg, Denmark. lisbeth.mikkelsen@plbio.kvl.dk
Sarrocco
Sabrina
S
Lübeck
Mette
M
Jensen
Dan Funck
DF
D015966:Gene Expression Regulation, Fungal; D020462:Gliocladium / Q000235:genetics; D049452:Green Fluorescent Proteins; D007202:Indicators and Reagents / Q000378:metabolism; D008164:Luminescent Proteins / Q000235:genetics*; D003904:Mitosporic Fungi / Q000235:genetics*; D010407:Penicillium / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D014242:Trichoderma / Q000235:genetics
D007202:Indicators and Reagents; D008164:Luminescent Proteins; C413663:fluorescent protein 583; D049452:Green Fluorescent Proteins