Bates G W
Department of Biological Science, Florida State University, Tallahassee 32306.
Mol Biotechnol. 1994 Oct;2(2):135-45. doi: 10.1007/BF02824806.
This article describes an optimized protocol for the electroporation of tobacco mesophyll protoplasts together with notes and data on the effects of various parameters and suggestions for work with protoplasts of other species. In this protocol, electroporation is achieved by means of electrical pulses from a high-voltage, capacitive-discharge unit. Procedures are described for measurement of protoplast viability with Evan's blue, the detection of transient expression of CAT and GUS gene plasmid constructs, and for the recovery of stable transformants based on selection for kanamycin resistance.
本文描述了一种优化的烟草叶肉原生质体电穿孔方案，以及关于各种参数影响的注释和数据，还有针对其他物种原生质体操作的建议。在该方案中，电穿孔通过高压电容放电装置产生的电脉冲来实现。文中描述了用伊文思蓝测定原生质体活力、检测CAT和GUS基因质粒构建体瞬时表达以及基于卡那霉素抗性选择来获得稳定转化体的步骤。
