A large-scale Agrobacterium-mediated transformation procedure with a strong positive-negative selection for gene targeting in rice (Oryza sativa L.)
A large-scale Agrobacterium-mediated transformation procedure with a strong positive-negative selection for gene targeting in rice (Oryza sativa L.)
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DOI：
10.1007/s00299-003-0752-0
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发表时间：
2004-04-01
影响因子：
6.2
通讯作者：
Iida, S
中科院分区：
文献类型：
--
作者：
Terada, R;Asao, H;Iida, S
用高效的阳性-阴性选择处理足够数量的稳定转化体的大规模转化程序是通过同源重组成功靶向基因的必要先决条件，因为据报道，与通过非同源末端连接的随机整合相比，通过体细胞同源重组在高等植物中的转基因整合为10（-3）至10（-5）。我们建立了一个有效的和大规模的农杆菌介导的水稻转化方案，产生约10（3）稳定的转化体常规从150种子和一个强大的正负选择程序，导致幸存者在10（-2）使用白喉毒素A片段的基因作为阴性标记。建立的转化程序为水稻基因打靶提供了基础。
A large-scale transformation procedure handling an adequate number of stable transformants with highly efficient positive-negative selection is a necessary prerequisite to successful gene targeting by homologous recombination, as the integration of a transgene by somatic homologous recombination in higher plants has been reported to be 10(-3) to 10(-5) compared with random integration by non-homologous end joining. We established an efficient and large-scale Agrobacterium-mediated rice transformation protocol that generated around 10(3) stable transformants routinely from 150 seeds and a strong positive-negative selection procedure that resulted in survivors at 10(-2) using the gene for diphtheria toxin A fragment as a negative marker. The established transformation procedure provides a basis for efficient gene targeting in rice.