摘要
利用含有四环素阻遏蛋白,具有卡娜霉素抗性的烟草种子为实验材料,采用根癌农杆菌介导转化中的叶圆盘法,将外源基因导入烟草愈伤组织,建成可诱导表达细胞质和细胞核CaM/CaM结合多肽的转基因烟草体系.在获得转基因植物的过程中,通过比较影响根癌农杆菌转化频率的各种因素,表明植物材料的选择和培养,农杆菌的培养和稀释,植物材料与农杆菌共培养的程度及转基因植株的筛选条件是影响农杆菌转化效率的重要因素.
The gene for CaM and CaMBp was transferred into tobacco leaf disks with help of Tiplasmid of Agrobacterium tumefactions.Tobacco leaf disks were rooted in MS medium containing hygromycin after preculture,agroinfection and coculture in 2～3 months.We established system of trangenic tobacco which will express calmodulin or calmodulinbinding protein.In this experiment, it was found there were many effect foctors.The effect of agroinfection on plant in different growth period was different and time of transgene were affected by different lines. The culture and dilution of Agrobacterium tumefactions influenced trangenic efficiency.The coculture of plant and Agrobacterium tumefactions aslo influenced trangenic efficiency.It was important for root of seedling to be screened that could select out transgene tobacco.
出处
《云南大学学报（自然科学版）》
CAS
CSCD
2003年第2期148-152,共5页
Journal of Yunnan University(Natural Sciences Edition)
基金
国家自然科学基金资助项目(39660009)
云南省自然科学基金资助项目(96CO42M
98CO006G
98CO0027).
关键词
烟草
根癌农杆菌
转基因
tobacco
Agrobacterium tumefactions
transgene
