Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 1565-1572 |
| Number of pages | 8 |
| Journal | Nature Protocols |
| Volume | 2 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - 2007 Jul 1 |
| Externally published | Yes |
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ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
Cite this
Arabidopsis mesophyll protoplasts : A versatile cell system for transient gene expression analysis. / Yoo, Sang-Dong; Cho, Young Hee; Sheen, Jen.
In: Nature Protocols, Vol. 2, No. 7, 01.07.2007, p. 1565-1572.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Arabidopsis mesophyll protoplasts
T2 - A versatile cell system for transient gene expression analysis
AU - Yoo, Sang-Dong
AU - Cho, Young Hee
AU - Sheen, Jen
PY - 2007/7/1
Y1 - 2007/7/1
N2 - 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.
AB - 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.
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U2 - 10.1038/nprot.2007.199
DO - 10.1038/nprot.2007.199
M3 - Article
VL - 2
SP - 1565
EP - 1572
JO - Nature Protocols
JF - Nature Protocols
SN - 1754-2189
IS - 7
ER -