Genetic introduction of foreign genes to Pleurotus eryngii by restriction enzyme-mediated integration.
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 microg of the HindIII-digested plasmid DNA and 10(6) 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.
Noh
Won
W
Department of Microbiology and Research Institute of Life Sciences, Gyeongsang National University, Chinju, 660-701, Republic of Korea.
Kim
Sang-Woo
SW
Dong-Won
Bae
B
Kim
Jae-Yean
JY
Ro
Hyeon-Su
HS
D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D005822:Genetic Vectors; D005827:Genetics, Microbial / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D010957:Plasmids; D020076:Pleurotus / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic
D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins