EXPRESSION OF GENES TRANSFERRED INTO MONOCOT AND DICOT PLANT-CELLS BY ELECTROPORATION
EXPRESSION OF GENES TRANSFERRED INTO MONOCOT AND DICOT PLANT-CELLS BY ELECTROPORATION
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DOI：
10.1073/pnas.82.17.5824
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发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
WALBOT, V
中科院分区：
文献类型：
--
作者：
FROMM, M;TAYLOR, LP;WALBOT, V
我们已经开发了一种将DNA电导入植物细胞的通用方法。当高压电脉冲施加到含有原生质体和DNA的溶液时，基因转移发生。以胡萝卜原生质体为模型系统，优化基因转移效率，电穿孔后24-48小时，通过引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。基因转移效率随着DNA浓度的增加而增加，并且受到电脉冲的幅度和持续时间以及电穿孔介质的组成的影响。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，表明该方法适用于单子叶植物和双子叶植物原生质体。
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 containting 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 introduce 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.