摘要
将含有大约50kb水稻基因组片段的TAC17克隆(NK15)通过电击转化到农杆菌LBA4404中,经多次继代培养,该克隆在农杆菌中是稳定的。用常规的农杆菌介导方法将该克隆转化粳稻品种农垦58S成熟胚的愈伤组织,对T0代进行PCR和Southern杂交分析表明,TAC17所携带的50kb外源DNA片段已完整地整合到水稻基因组上,整合方式多数为单位点插入,整合位点是随机的。经T1代分析表明,外源基因可以稳定地遗传,而且进一步确定外源大片段的整合方式为为单位点插入。
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 T_1 progeny confirmed that the inserted forgein DNA was stably inherited.
基金
国家重点基础研究发展规划(973项目)(编号:G1999011603)
国家"863"计划(编号:2002AA224121)
国家自然科学基金(编号: 30080019)项目资助~~
