拟南芥原生质体瞬时表达体系的改进

Q1 Agricultural and Biological Sciences

Current protocols in plant biology Pub Date : 2016-06-10 DOI:10.1002/cppb.20013

Yangrong Cao, Hong Li, An Q. Pham, Gary Stacey

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引用次数: 11

摘要

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、蛋白质关联和基因表达调控等提供了有力的工具。有几种方法，包括电穿孔，已报道将DNA引入原生质体。然而，使用的最佳方法之一是聚乙二醇(PEG)介导的转染。在这里，我们描述了一种改进的peg介导转化方法，包括原生质体的制备，peg介导的转化，以及AtLYK5基因(AT2G33580)在原生质体中的表达。原生质体瞬时表达系统为植物生物化学和生理学的细胞实验提供了独特的支持能力。©2016 by John Wiley &儿子,Inc。

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An Improved Transient Expression System Using Arabidopsis Protoplasts

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.

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来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

自引率

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发文量

期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.