电击介导的细胞转染。电学参数的表征与优化。
Electric shock-mediated transfection of cells. Characterization and optimization of electrical parameters.
作者信息
Winterbourne D J, Thomas S, Hermon-Taylor J, Hussain I, Johnstone A P
机构信息
Department of Surgery, St George's Hospital Medical School, London, U.K.
出版信息
Biochem J. 1988 Apr 15;251(2):427-34. doi: 10.1042/bj2510427.
Abstract
The effect of various parameters on the electric shock-mediated permeabilization and transfection of CHO cells has been investigated. Up to 70% of the cells can be maintained transiently permeable to erythrosin B for periods of at least 1 h at 20 degrees C. Electrical conditions optimal for transient permeabilization were also optimal for efficient DNA transfection by pSV2neo. However, the DNA must be present during exposure to the electric field for efficient transformation. The same requirement existed for voltage-induced DNA toxicity. The results suggest that DNA moves into the cells by electrophoresis, not by simple diffusion. Based on these observations a simple, rapid procedure for optimizing the conditions for electric shock-mediated DNA transfer into cells has been developed.
摘要
研究了各种参数对电休克介导的中国仓鼠卵巢细胞（CHO细胞）通透性和转染的影响。在20℃下，高达70%的细胞可在至少1小时内保持对赤藓红B的瞬时通透性。瞬时通透化的最佳电条件也是通过pSV2neo进行高效DNA转染的最佳条件。然而，为了实现高效转化，DNA必须在暴露于电场期间存在。电压诱导的DNA毒性也存在同样的要求。结果表明，DNA通过电泳进入细胞，而不是通过简单扩散。基于这些观察结果，开发了一种简单、快速的程序来优化电休克介导的DNA转入细胞的条件。
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