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This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

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Lignin is an obstacle in large-scale biofuel production. This can be overcome by the use of fungal pretreatment. This experiment assessed two transformation assays, based on the protocols of Laccaria bicolor and Phanerochaete chrysosporium, and applied them to Trametes versicolor. In order to determine a successful transformation, T. versicolor was transformed with green fluorescence protein (GFP) using the two assays, followed by detection of transformed mycelia under UV light. The two protocols were assessed based on timeliness, ease, and effectiveness. Based on the quantity of viable transformants and ease, the membrane assay described for Laccaria is successful when the antibiotic concentration, specifically, is optimized and applied to T. versicolor. This work is also the first to show that the plant-based promoter, cauliflower mosaic virus (CaMV) 35S, is effective for expression of any gene in T. versicolor, a white-rot fungus.

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