Cao Yangrong, Li Hong, Pham An Q, Stacey Gary
State Key Lab of Agricultural Microbiology, College of Life Science Technology, Huazhong Agricultural University, Wuhan, Hubei, China.
Division of Plant Science, National Center for Soybean Biotechnology, University of Missouri, Columbia, Missouri.
Curr Protoc Plant Biol. 2016 Mar;1(2):285-291. doi: 10.1002/cppb.20013.
Transient gene expression in protoplasts provides a powerful tool to study protein expression, protein localization, protein-protein association, and gene expression regulation, etc. There are several methods including electroporation, which have been reported to introduce DNA into protoplasts. However, one of the best methods used is polyethylene glycol (PEG)-mediated transfection. Here, we describe an improved PEG-mediated transformation method including preparation of protoplasts, PEG-mediated transformation, and, by way of example, expression of the AtLYK5 gene (AT2G33580) in protoplasts. The protoplast transient expression system provides unique capabilities to support cell-based experiments involved in plant biochemistry and physiology. © 2016 by John Wiley & Sons, Inc.
原生质体中的瞬时基因表达为研究蛋白质表达、蛋白质定位、蛋白质-蛋白质相互作用以及基因表达调控等提供了一个强大的工具。已有多种方法（包括电穿孔法）被报道可将DNA导入原生质体。然而，常用的最佳方法之一是聚乙二醇（PEG）介导的转染。在此，我们描述一种改进的PEG介导的转化方法，包括原生质体的制备、PEG介导的转化，并举例说明AtLYK5基因（AT2G33580）在原生质体中的表达。原生质体瞬时表达系统为支持植物生物化学和生理学中基于细胞的实验提供了独特的能力。© 2016约翰威立国际出版公司
