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pGREEN is a flexible Ti binary vector for Agrobacterium mediated transformation system . The advantage of this vector is that optional combinations of selection and reporter gene cassettes can be optimally assembled to meet different transformation requirements. gfp (green fluorescent protein) gene driven by 35S-CaMV promoter was used as a reporter gene to enable transformant detection in vivo and the REDExtract-N-Amp Plant PCR Kit was applied as a high throughput approach for screening large amount of regenerated transformed plants in this report. With the designated vector pGK assembled using pGreen0029 and 35S-GFP, different phenotypes of transformation chimerism were observed and average transformation rates 7.75% and 2.03 % were acquired in term of consistent PCR results between the selectable and the reporter gene for empty vector and inserted recombinants respectively in transformation events of citrus grapefruits.
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Author(s): Qifa Zheng Chunxian Chen Shu Huang Xu Xiang Young A Choi Fred G Gmitter Jr
Agrobacterium, transformation, citrus, GFP (green fluorescent protein), PCR