将重组质粒高效导入农杆菌的电脉冲转化研究  

Efficient transformation of recombinant plasmid DNA by electroporation into Agrobacterium tumefaciens

作　　者：杨明[1] 黄兴奇[1] 张绍松[1] 万萌[1] 鄢波[1] 

机构地区：[1]云南省农业科学院,中国昆明650205

出　　处：《西南农业学报》2000年第2期1-4,共4页Southwest China Journal of Agricultural Sciences

摘　　要：利用电脉冲法将重组质粒ＰＢＩ１２１／α—ＰＡＰ导入农杆菌ＬＢＡ４４０４，获得 ３．５×１０６转化子／μ ｇ质粒 ＤＮＡ的转化效率。探讨了不同电激条件及农杆菌细胞生 长状态对转化率的影响。结果表明，农杆菌ＬＢＡ４４０４电脉冲转化的最适电激条件 为电场强度 １８ｋｖ／ｃｍ，电容 ２５μＦ，电阻 ２００Ω；农杆菌最适细胞生长状态为 ＯＤ６００ ＝０．４～０．５。Efficient transformation of Agrobacterium tumefaciens LBA4404 by electroporation was performed using the binary vector PAI121 that contain α-PAP gene and 35s promoter. The optimal electroporation condition for plasmid transfer was obtained at 18kv/cm, 25μF and 200Ω. And the growth stage of A. tumefaciens was OD600= 0. 4 --0. 5. The frequency of transformation was a linear function of the DNA concentration.

关 键 词：农杆菌 电脉冲 电激法 高效转化 重组质粒 

分 类 号：S188[农业科学—农业基础科学] Q78[生物学—分子生物学]