[基于转化能人工染色体(TAC)载体的水稻转化体系的建立]。

Ling-Yan Zhou, Da-Gang Jiang, Hao Wu, Chu-Xiong Zhuang, Yao-Guang Liu, Man-Tong Mei
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引用次数: 0

摘要

采用电穿孔法将含有约50 kb DNA插入片段的TAC克隆(NK15)导入农杆菌LBA4404。在卡那霉素选择下,NK15在农杆菌LBA4404中稳定存在。用携带NK15的农垦58s成熟胚愈伤组织感染农垦58s成熟胚愈伤组织。转基因植株的PCR和Southern分析表明,这50 kb的外源DNA被转移到水稻基因组中,大多数转基因植株都有一个插入拷贝。对T1子代的遗传和PCR分析证实,插入的外源蛋白DNA是稳定遗传的。
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[Development of transformation system of rice based on transformation-competent artificial chromosome (TAC) vector].

The TAC clone (NK15) containing a ca.50 kb DNA insert was introduced into Agrobacterium tumefaciens strain LBA4404 by electroporation. The NK15 was stable in Agrobacterium tumefaciens strain LBA4404 under kanamycin selection for many generations. The calli of mature embryo of Nongken58S were infected with the Agrobacterium tumefaciens strain LBA4404 carrying NK15. PCR and Southern analyses of transgenic plants indicated that the 50 kb of foreign DNA was transferred into the rice genome, and most of transgenic plants had one copy of the insertion. Genetic and PCR analyses of T1 progeny confirmed that the inserted forgein DNA was stably inherited.

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