Fromm M, Taylor L P, Walbot V
Proc Natl Acad Sci U S A. 1985 Sep;82(17):5824-8. doi: 10.1073/pnas.82.17.5824.
We have developed a general method for electrically introducing DNA into plant cells. Gene transfer occurs when a high-voltage electric pulse is applied to a solution containing protoplasts and DNA. Carrot protoplasts were used as a model system to optimize gene-transfer efficiency, which was measured 24-48 hr after electroporation by the amount of chloramphenicol acetyltransferase activity resulting from the expression of the introduced chimeric plasmids. Gene-transfer efficiency increased with the DNA concentration and was affected by the amplitude and duration of the electric pulse as well as by the composition of the electroporation medium. Our optimized gene-transfer conditions were effective when applied to tobacco and maize protoplasts, demonstrating that the method is applicable to both monocot and dicot protoplasts.
我们已经开发出一种将DNA电导入植物细胞的通用方法。当向含有原生质体和DNA的溶液施加高压电脉冲时，基因转移就会发生。胡萝卜原生质体被用作模型系统来优化基因转移效率，在电穿孔后24 - 48小时，通过导入的嵌合质粒表达产生的氯霉素乙酰转移酶活性的量来测量基因转移效率。基因转移效率随DNA浓度的增加而提高，并受电脉冲的幅度和持续时间以及电穿孔介质组成的影响。我们优化的基因转移条件应用于烟草和玉米原生质体时是有效的，这表明该方法适用于单子叶和双子叶原生质体。
