Zhou Ling-Yan, Jiang Da-Gang, Wu Hao, Zhuang Chu-Xiong, Liu Yao-Guang, Mei Man-Tong
Genetic Engineeering Laboratory, College of Life Science, South China Agricutural University, Guangzhou 510642, China.
Yi Chuan Xue Bao. 2005 May;32(5):514-8.
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.
将含有约50 kb DNA插入片段的TAC克隆(NK15)通过电穿孔法导入根癌农杆菌菌株LBA4404。在卡那霉素选择下,NK15在根癌农杆菌菌株LBA4404中稳定存在许多代。用携带NK15的根癌农杆菌菌株LBA4404感染农垦58S成熟胚愈伤组织。对转基因植株的PCR和Southern分析表明,50 kb的外源DNA已转移到水稻基因组中,且大多数转基因植株有一个插入拷贝。T1代子代的遗传和PCR分析证实插入的外源DNA能稳定遗传。