Raptis L H, Liu S K, Firth K L, Stiles C D, Alberta J A
Department of Microbiology and Immunology, Queen's University, Kingston, Ontario, Canada.
Biotechniques. 1995 Jan;18(1):104, 106, 108, 110 passim.
To study the effect of protein interactions in vivo upon cellular functions,such complexes may be disrupted through the introduction of peptides corresponding to the proteins' points of contact. In this communication a simple, rapid and reproducible procedure for peptide introduction into adherent cells by electroporation is described. Cells are grown on electrically conductive, optically transparent indium-tin oxide at the time of pulse delivery. Several electrode and slide configurations, necessary to obtain non-electroporated cells adjacent to the electroporated ones as a control, are outlined. Careful control of electric field strength achieved the introduction of the peptide into essentially 100% of the cells while this treatment caused no detectable disruption of their division cycle.
为了研究体内蛋白质相互作用对细胞功能的影响,可以通过引入与蛋白质接触点对应的肽来破坏此类复合物。在本通讯中,描述了一种通过电穿孔将肽引入贴壁细胞的简单、快速且可重复的方法。在施加脉冲时,细胞生长在导电的、光学透明的氧化铟锡上。概述了几种电极和载玻片配置,这些配置对于获得与电穿孔细胞相邻的未电穿孔细胞作为对照是必要的。对电场强度的仔细控制实现了将肽引入基本上100%的细胞中,同时这种处理未导致其分裂周期出现可检测到的破坏。