HIGH-EFFICIENCY TRANSFORMATION OF BACTERIAL-CELLS BY ELECTROPORATION
HIGH-EFFICIENCY TRANSFORMATION OF BACTERIAL-CELLS BY ELECTROPORATION
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DOI：
10.1128/jb.170.6.2796-2801.1988
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发表时间：
1988-06-01
影响因子：
3.2
通讯作者：
HANAWALT, PC
中科院分区：
文献类型：
--
作者：
CALVIN, NM;HANAWALT, PC
我们已经开发了一种方法，通过使用一种新型的电穿孔装置，在大肠杆菌K-12衍生物的外膜中有效地产生瞬时孔。这些孔足够大并且持续足够长的时间以促进质粒分子在细胞内和细胞外空间之间的平衡。该方法已用于转化细菌细胞，转化效率大于109个转化体/μ g质粒。它也被用来提取完整的质粒从转化细胞的效率与传统的碱裂解或氯化铯平衡密度梯度技术。该技术简单快速，允许在几分钟内完成微克量质粒的转化或制备。
We have developed a method for efficiently generating transient pores in the outer membranes of Escherichia coli K-12 derivatives by using a new type of electroporation apparatus. The pores are large enough and persist long enough to facilitate the equilibration of plasmid molecules between the intracellular and extracellular spaces. The method has been used to transform bacterial cells with an efficiency greater than 109 transformants per .mu.g of plasmid. It has also been used to extract intact plasmid from transformed cells with efficiencies comparable to those of the traditional alkaline lysis or CsCl equilibrium density gradient techniques. The technique is simple and rapid, allowing a transformation or the preparation of microgram quantities of plasmid to be accomplished in minutes.