Arabidopsis mesophyll protoplasts:: a versatile cell system for transient gene expression analysis

被引：4042

作者：

Yoo, Sang-Dong ^{[1
,2
]}

Cho, Young-Hee ^{[1
,2
]}

Sheen, Jen ^{[1
,2
]}

机构：

[1] Massachusetts Gen Hosp, Dept Mol Biol, Boston, MA 02114 USA

[2] Harvard Univ, Sch Med, Dept Genet, Boston, MA 02114 USA

来源：

NATURE PROTOCOLS | 2007年 / 2卷 / 07期

关键词：

D O I：

10.1038/nprot.2007.199

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The transient gene expression system using Arabidopsis mesophyll protoplasts has proven an important and versatile tool for conducting cell-based experiments using molecular, cellular, biochemical, genetic, genomic and proteomic approaches to analyze the functions of diverse signaling pathways and cellular machineries. A well-established protocol that has been extensively tested and applied in numerous experiments is presented here. The method includes protoplast isolation, PEG-calcium transfection of plasmid DNA and protoplast culture. Physiological responses and high-throughput capability enable facile and cost-effective explorations as well as hypothesis-driven tests. The protoplast isolation and DNA transfection procedures take 6-8 h, and the results can be obtained in 2-24 h. The cell system offers reliable guidelines for further comprehensive analysis of complex regulatory mechanisms in whole-plant physiology, immunity, growth and development.

引用

页码：1565 / 1572

页数：8

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