Transformation of Lithium Acetate-treated Neurospora crassa
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DNA-mediated transformation provides a powerful tool for both genome analysis and gene manipulation. Advances in transformation procedures of filamentous fungi have played a key role in making these organisms tractable on a molecular genetic level. A number of different transformation protocols exist for fungal transformation, each with inherent advantages for different applications. To be described here, lithium acetate has been used with success in preparing Neurospora crassa conidia for transformation. The technique involves the treatment of germinating conidia consecutively with (1) lithium acetate, (2) transforming DNA, and (3) PEG; followed by a brief heat shock before plating. The lithium acetate procedure provides a means for rapid and efficient transformation of N. crassa when moderate transformation frequencies are useful. Further, the technique has general applicability to a variety of fungal species.
KeywordsNeurospora crassa Lithium acetate Transforming DNA PEG Filamentous fungi Exogenous DNA
This research was supported by a Medical Innovations Grant from the Wright State University Boonshoft School of Medicine. I thank Heather Hostetler for microscopy assistance.
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