Abstract
Pleurotus eryngii was transformed using a polyethylene glycol-mediated method. A plasmid, pEPUGH, containing a reporter gene (enhanced green fluorescent protein gene, egfp) and a positive selectable marker gene (hygromycin phosphotransferase gene, hph) was constructed. The fused egfp-hph gene was placed under the control of the strong and constitutive native gpd promoter from P. eryngii. The recombinant plasmid was used to transform of P. eryngii protoplasts. Successful transformation was demonstrated by molecular analyses. Moreover, the mycelia of the transformants showed green epipolic dispersion on fluorescence microscopy. About 90-210 transformants were produced per μg plasmid DNA per 10(7) viable protoplasts.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Genes, Reporter
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Glyceraldehyde-3-Phosphate Dehydrogenases / genetics
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Green Fluorescent Proteins / chemistry
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Green Fluorescent Proteins / genetics*
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Green Fluorescent Proteins / metabolism
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Mycelium / growth & development
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Phosphotransferases (Alcohol Group Acceptor) / genetics
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Plasmids / genetics
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Pleurotus / chemistry
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Pleurotus / genetics*
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Pleurotus / metabolism
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Polyethylene Glycols / chemistry
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Polyethylene Glycols / metabolism*
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Promoter Regions, Genetic
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Real-Time Polymerase Chain Reaction
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics*
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Recombinant Fusion Proteins / metabolism
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Transfection / methods*
Substances
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Recombinant Fusion Proteins
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enhanced green fluorescent protein
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Green Fluorescent Proteins
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Polyethylene Glycols
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Glyceraldehyde-3-Phosphate Dehydrogenases
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Phosphotransferases (Alcohol Group Acceptor)
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hygromycin-B kinase