Development of ToxA and ToxB promoter-driven fluorescent protein expression vectors for use in filamentous ascomycetes.
The green fluorescent protein (GFP) has been established as the premier in vivo reporter for investigations of gene expression, protein localization, and cell and organism dynamics. The fungal transformation vector pCT74, with sGFP under the control of the ToxA promoter from Pyrenophora tritici-repentis, effectively expresses GFP in a diverse group of filamentous ascomycetes. Due to the versatility of ToxA promoter-driven expression of GFP, we constructed an additional set of fluorescent protein expression vectors to expand the color palette of fluorescent markers for use in filamentous fungi. EYFP, ECFP and mRFP1 were successfully expressed from the ToxA promoter in its fungus of origin, P. tritici-repentis, and a distant relative, Verticillium dahliae. Additionally the ToxB promoter from P. tritici-repentis drove expression of sGFP in V. dahliae, suggesting a similar potential to the ToxA promoter for heterologous expression in ascomycetes. The suite of fungal transformation vectors presented here promise to be useful for a variety of fungal research applications.
Andrie
Rachael M
RM
Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331, USA.
Martinez
J Patrick
JP
Ciuffetti
Lynda M
LM
D001203:Ascomycota / Q000235:genetics*; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D017930:Genes, Reporter; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D008856:Microscopy, Fluorescence; D008967:Molecular Biology / Q000379:methods; D009183:Mycotoxins / Q000235:genetics*; D011401:Promoter Regions, Genetic; D000097573:Red Fluorescent Protein
D001426:Bacterial Proteins; D005656:Fungal Proteins; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D009183:Mycotoxins; D000097573:Red Fluorescent Protein; C485182:Cyan Fluorescent Protein; C105776:TOXA protein, Pyrenophora tritici-repentis; C434209:ToxB protein, Pyrenophora tritici-repentis; C054562:yellow fluorescent protein, Bacteria