Potter Huntington, Heller Richard
Byrd Alzheimer's Institute, University of South Florida College of Medicine, Tampa, Florida, USA.
Curr Protoc Mol Biol. 2010 Oct;Chapter 9:Unit9.3. doi: 10.1002/0471142727.mb0903s92.
Electroporation--the use of high-voltage electric shocks to introduce DNA into cells--can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit describes electroporation of mammalian cells, including ES cells for the preparation of knock-out, knock-in, and transgenic mice. Protocols are described for the use of electroporation in vivo to perform gene therapy for cancer therapy and DNA vaccination. Also described are modifications for preparation and transfection of plant protoplasts.
电穿孔法——利用高压电击将DNA导入细胞——可用于大多数细胞类型，能产生高频率的稳定转化和瞬时基因表达，而且由于所需步骤较少，可能比其他技术更简便。本单元介绍了哺乳动物细胞的电穿孔法，包括用于制备基因敲除、基因敲入和转基因小鼠的胚胎干细胞。还描述了体内电穿孔用于癌症治疗的基因治疗和DNA疫苗接种的方案。此外，还介绍了植物原生质体制备和转染的改良方法。
