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Development of Marker-free Transgenic Rice (Oryza Sativa L.) Via Agrobacterium- Mediated Method with cre/loxP-argE System KCI 등재
- 언어ENG
- URLhttps://db.koreascholar.com/Article/Detail/319695
Until now many strategies have been used to produce marker-free transgenic plants such as co-transformation with negative selectable markers, site-specific recombination system, transposable elements mediated transformation, and etc. In this research, embryogenic calli induced from japonica rices, Ilmibyeo and Dongjinbyeo, were tranformed with the vector which simultaneously constructed with cre/loxP and argE genes in T-DNA. Transformation efficiencies were comparably lower than those of our previous studies, since the constructed genome size was relatively big (>10Kb). For eliminate the transformed tissues which contained positive selectable marker gene, tunicamycin was treated at regeneration and selection stages, since cre recombinase gene is expressed under the presence of this antibiotics. The plants were selected first under 50 mg․L-1 hygromycin at 28℃ for 2 weeks after the Agrobacterium-infection at 25℃ for 7 days. And then, the regeneration plants were successfully obtained on MS basal regeneration medium containing 0.1 mg․L-1 tunicamycin. The regenerated plants are now acclimatizing in greenhouse and molecular analysis are currently accomplished with these plants.
