电转化法提高平连载体DNA转化效率的研究

IMPROVING TRANSFORMATION EFFICIENCY OFBLUNT-END LIGATED VECTOR DNA BY ELECTROTRANSFORMATION

探讨了电转化过程中一些影响平连载体ＤＮＡ转化效率的因素，并与化学法进行了比较，由此建立了优化的电转化条件。结果显示，在ＯＤ６００值０．７２－０．７８收获细胞时，可使平连载体ＤＮＡ的转化效率达到５×１０６转化子／μｇｃＤ－ＮＡ，较化学法高１０２－１０３倍。同时，降低连接产物的盐浓度。

A number of factors that infected transformation efficiency of blunt-end ligated vector were investigated in this paper. By contrasting transformation efficiency with chemical, an improved electrotransformation protocol was establisheed. The results showed that the transformation efficiency of blunt-end ligated vector can be up to 5x10 6 transformants from 1 μg cDNA when cells were harvested in OD 600 0. 72-0.78, which was in a 10 2-10 3 fold increase by comparing with chemical. In the same time, it was important for the success of electrotransformation and the improvement of transformation efficiency to lower salt concentration of ligation products.