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KMID : 0880220100480020253
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Journal of Microbiology
2010 Volume.48 No. 2 p.253 ~ p.256
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Genetic Introduction of Foreign Genes to Pleurotus eryngii by Restriction Enzyme-Mediated Integration
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Noh Won
Kim Sang-Woo
Bae Dong-Won
Kim Jae-Yean
Ro Hyeon-Su
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Abstract
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Pleurotus eryngii was transformed via restriction enzyme-mediated integration. In order to construct the transformation plasmid, the enhanced cyan fluorescent protein (ECFP) gene was ligated next to the gpd promoter of the plasmid pAN7-1. Transformation was facilitated via the heat treatment of a transformation mixture containing 1 μg of the HindIII-digested plasmid DNA and 106 mushroom protoplasts in 40% polyethyleneglycol solution, resulting in 10-40 hygromycin-resistant transformants. Successful transformation was evidenced by PCR, Southern blot, and confocal fluorescence microscopic analyses on the selected transformants. To date, this is the first report on the transformation of P. eryngii by REMI technique.
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KEYWORD
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hygromycin, P. eryngii, REMI, transformation
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