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An improved protocol for<Emphasis Type="Italic"> Agrobacterium</Emphasis>-mediated transformation of<Emphasis Type="Italic"> Antirrhinum majus</Emphasis> L.
Authors:M-L?Cui  T?Handa  Email author" target="_blank">H?EzuraEmail author
Institution:(1) Gene Research Center, University of Tsukuba, 305 8572 Tsukuba, Ibaraki, Japan;(2) Institute of Agriculture and Forestry, University of Tsukuba, 305 8572 Tsukuba, Ibaraki, Japan
Abstract:Efficient Agrobacterium -mediated transformation of Antirrhinum majus L. was achieved via indirect shoot organogenesis from hypocotyl explants of seedlings. Stable transformants were obtained by inoculating explants with A. tumefaciens strain GV2260 harboring the binary vector pBIGFP121, which contains the neomycin phosphotransferase gene (NPT II) as a selectable marker and the gene for the Green Fluorescent Protein (GFP) as a visual marker. Putative transformants were identified by selection for kanamycin resistance and by examining the shoots using fluorescence microscopy. PCR and Southern analyses confirmed integration of the GFP gene into the genomes of the transformants. The transformants had a morphologically normal phenotype. The transgene was shown to be inherited in a Mendelian manner. This improved method requires only a small number of seeds for explant preparation, and three changes of medium; the overall transformation efficiency achieved, based on the recovery of transformed plants after 4–5 months of culture, reached 8–9%. This success rate makes the protocol very useful for producing transgenic A. majus plants.Communicated by G. Jürgens
Keywords: Antirrhinum majus L    Green fluorescent protein gene (GFP)  Hypocotyl explants   Agrobacterium-mediated transformation
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