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- Publisher Website: 10.1007/s00299-010-0953-2
- Scopus: eid_2-s2.0-79952699240
- PMID: 21103876
- WOS: WOS:000288396800003
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Article: Protoplast isolation and transient gene expression in the single-cell C4 species, Bienertia sinuspersici
Title | Protoplast isolation and transient gene expression in the single-cell C<inf>4</inf> species, Bienertia sinuspersici |
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Authors | |
Keywords | Bienertia sinuspersici Chlorenchyma cell Photosynthesis Protoplast Single-cell C 4 Transient expression |
Issue Date | 2011 |
Citation | Plant Cell Reports, 2011, v. 30, n. 4, p. 473-484 How to Cite? |
Abstract | Although transient gene expression using reporters such as green fluorescent protein is a versatile tool for examining gene functions and intracellular protein trafficking, the establishment of a highly efficient gene manipulation method remains a challenge in many plant species. A reliable transformation protocol has not yet been established for the three single-cell C4 species, despite their potential of serving as model systems for their extraordinary C4 photosynthetic metabolism. We report the first protocol optimized for isolating a large-scale and homogenous population of protoplasts from chlorenchyma cells of the single-cell C4 species Bienertia sinuspersici. Cytochemical staining confirmed the preservation of the unusual subcellular compartmentation of organelles in chlorenchyma cells after cell wall digestion. Approximately 84% of isolated protoplasts expressed the reporter fluorescent protein following our optimized polyethylene glycol-mediated transfection procedures. Fluorescent fusion protein tagged with various intracellular sorting signals demonstrated potential use of the transient gene expression system in subcellular protein localization and organelle dynamics studies. Further applications of the current protoplast isolation and transfection techniques in understanding the novel single-cell C4 photosynthetic mechanism are discussed. © 2010 Springer-Verlag. |
Persistent Identifier | http://hdl.handle.net/10722/311368 |
ISSN | 2023 Impact Factor: 5.3 2023 SCImago Journal Rankings: 1.319 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Lung, Shiu Cheung | - |
dc.contributor.author | Yanagisawa, Makoto | - |
dc.contributor.author | Chuong, Simon D.X. | - |
dc.date.accessioned | 2022-03-22T11:53:46Z | - |
dc.date.available | 2022-03-22T11:53:46Z | - |
dc.date.issued | 2011 | - |
dc.identifier.citation | Plant Cell Reports, 2011, v. 30, n. 4, p. 473-484 | - |
dc.identifier.issn | 0721-7714 | - |
dc.identifier.uri | http://hdl.handle.net/10722/311368 | - |
dc.description.abstract | Although transient gene expression using reporters such as green fluorescent protein is a versatile tool for examining gene functions and intracellular protein trafficking, the establishment of a highly efficient gene manipulation method remains a challenge in many plant species. A reliable transformation protocol has not yet been established for the three single-cell C4 species, despite their potential of serving as model systems for their extraordinary C4 photosynthetic metabolism. We report the first protocol optimized for isolating a large-scale and homogenous population of protoplasts from chlorenchyma cells of the single-cell C4 species Bienertia sinuspersici. Cytochemical staining confirmed the preservation of the unusual subcellular compartmentation of organelles in chlorenchyma cells after cell wall digestion. Approximately 84% of isolated protoplasts expressed the reporter fluorescent protein following our optimized polyethylene glycol-mediated transfection procedures. Fluorescent fusion protein tagged with various intracellular sorting signals demonstrated potential use of the transient gene expression system in subcellular protein localization and organelle dynamics studies. Further applications of the current protoplast isolation and transfection techniques in understanding the novel single-cell C4 photosynthetic mechanism are discussed. © 2010 Springer-Verlag. | - |
dc.language | eng | - |
dc.relation.ispartof | Plant Cell Reports | - |
dc.subject | Bienertia sinuspersici | - |
dc.subject | Chlorenchyma cell | - |
dc.subject | Photosynthesis | - |
dc.subject | Protoplast | - |
dc.subject | Single-cell C 4 | - |
dc.subject | Transient expression | - |
dc.title | Protoplast isolation and transient gene expression in the single-cell C<inf>4</inf> species, Bienertia sinuspersici | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1007/s00299-010-0953-2 | - |
dc.identifier.pmid | 21103876 | - |
dc.identifier.scopus | eid_2-s2.0-79952699240 | - |
dc.identifier.volume | 30 | - |
dc.identifier.issue | 4 | - |
dc.identifier.spage | 473 | - |
dc.identifier.epage | 484 | - |
dc.identifier.isi | WOS:000288396800003 | - |