Development of ToxA and ToxB promoter-driven fluorescent protein expression vectors for use in filamentous ascomycetes

• Published in: Mycologia Vol. 97; no. 5; pp. 1152 - 1161
• Main Authors: Andrie, Rachael M., Martinez, J. Patrick, Ciuffetti, Lynda M.
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01-09-2005; Mycological Society of America
• Subjects: Ascomycetes; Ascomycota; Ascomycota - genetics; Bacterial Proteins - biosynthesis; Bacterial Proteins - genetics; Conidia; Fluorescence; fluorescent proteins; fungal promoter; Fungal Proteins - genetics; Fungi; gene expression regulation; Genes, Reporter; Genetic Vectors; green fluorescent protein; Green Fluorescent Proteins - biosynthesis; Green Fluorescent Proteins - genetics; heterologous expression; Imaging; Luminescent Proteins - biosynthesis; Luminescent Proteins - genetics; Microscopy; Microscopy, Fluorescence; Molecular Biology - methods; Mycotoxins - genetics; Polymerase chain reaction; promoter regions; Promoter Regions, Genetic; Protoplasts; Pyrenophora tritici-repentis; Red Fluorescent Protein; reporter genes; Techniques; ToxA; ToxB; Verticillium; Verticillium dahliae
• ISSN: 0027-5514; 1557-2536
• DOI: 10.1080/15572536.2006.11832763

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.