High efficiency of stable genetic transformation in Dendrobium via microprojectile bombardment
作者： C. Suwanaketchanatit , J. Piluek , S. Peyachoknagul , P. S. Huehne
DOI: 10.1007/S10535-007-0148-Z
关键词: Transformation (genetics) 、 Green fluorescent protein 、 Transformation efficiency 、 Plasmid 、 Southern blot 、 Dendrobium 、 Transgene 、 Molecular biology 、 genomic DNA 、 Biology
摘要: Several parameters affecting transient β-glucuronidase (GUS) expression in protocorms of Dendrobium cv. Jaquelyn Thomas were examined using bombardment technique with pActin-1D plasmid. The GUS activity protocorm was not significantly affected by size the target, type particles, and helium gas pressure. However, numbers surviving tissues after different. Transgenic orchids established actively dividing gold particles coated pMNK1005 plasmid containing hygromycin phosphotransferase (hgh) green fluorescent protein (gfp) genes driven a ubiquitin promoter. A high efficiency orchid transformation three selection steps. bombarded screened on medium supplemented 5 mg dm−3 25 stringently selected 30 hygromycin. 19.87 % GFP expressing chimeras. integration transgene into genomic DNA transgenic plantlets confirmed PCR Southern blot hybridization. All but one lines contained multiple copies transgene.
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