title: Polyethylene glycol-mediated transformation of fused egfp-hph gene under the control of gpd promoter in Pleurotus eryngii.
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.
authors: Institute of Plant and Environment Protection, Beijing Academy of Agriculture and Forestry Science, Beijing 100097, China.
authors: Yonggang
authors: Y
authors: Yin
authors: Yu
authors: Y
authors: Liu
authors: Haojie
authors: H
authors: Jin
authors: Shouxian
authors: S
authors: Wang
authors: Shuang
authors: S
authors: Zhao
authors: Xiaoli
authors: X
authors: Geng
authors: Ming
authors: M
authors: Li
authors: Feng
authors: F
authors: Xu
mesh_terms: D017930:Genes, Reporter; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D025282:Mycelium / Q000254:growth & development; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D020076:Pleurotus / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D060888:Real-Time Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014162:Transfection / Q000379:methods*
chemical_list: D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase