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Genetic introduction of foreign genes to <Emphasis Type="Italic">Pleurotus eryngii</Emphasis> by restriction enzyme-mediated integration
Authors:Won Noh  Sang-Woo Kim  Bae Dong-Won  Jae-Yean Kim  Hyeon-Su Ro
Institution:1.Department of Microbiology and Research Institute of Life Sciences,Gyeongsang National University,Chinju,Republic of Korea;2.Central Instrument Facility,Gyeongsang National University,Chinju,Republic of Korea;3.Division of Applied Life Science and Environmental Biotechnology National Core Research Center,Gyeongsang National University,Chinju,Republic of Korea
Abstract: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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