摘要
利用电激法将双元载体质粒ＰＢＩ１２１导入根癌农杆菌ＡＧＬ－１并获得较高转化效率．探讨了不同电激条件对转化效率的影响，并检测了ＣａＭＶ３５Ｓ启动子启动ＧＵＳ基因在根癌农杆菌中的表达．结果表明：根癌农杆菌电激转化的最适电激条件为１１ｋＶ／ｃｍ，２１．５μＦ，电场强度是影响转化效率的关键因素．当质粒ＤＮＡ浓度从０．２ｍｇ／Ｌ增加至０．６ｍｇ／Ｌ时，转化效率呈线性增加．细菌密度对转化效率也有一定影响．最高转化效率为每μｇ质粒ＤＮＡ１．７×１０５转化子．组织化学反应证实ＣａＭＶ３５Ｓ能启动ＧＵＳ基因在根癌农杆菌中表达．
Efficient transformation of Agrobacterium tumefaciens AGL-1 by electroporation was performed using the binary vector pBI 121 that contain β-glucuronidase(GUS)gene underthe control of the CaMV35S promoter.The optimal electroporatlon condition for plasmid transfer was obtained at 11 kV/cm and 21. 5μF,the field strength showed important influence on the transformation efficiency,The concentration of plasmid DNA and the density Of bacteria in the DNA/bacteria suspension also affected the transformation efficiency.Histochemical detection of GUS indicated that the transformants are capable of expressing GUS gene underthe control of the CaMV35S promoter.
出处
《武汉大学学报（自然科学版）》
CSCD
1995年第6期724-728,共5页
Journal of Wuhan University(Natural Science Edition)
基金
国家自然科学基金
关键词
转化
根癌农杆菌
电激法
农杆菌介导法
electroporation,transformation,Agrobacterium tumefaciens
