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Journal ArticleDOI

Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants.

Imogen Sparkes, +3 more
- 01 Jan 2006 - 
- Vol. 1, Iss: 4, pp 2019-2025
TLDR
The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway and is a relatively fast technique to assess expression of genes of interest.
Abstract
Expression and tracking of fluorescent fusion proteins has revolutionized our understanding of basic concepts in cell biology. The protocol presented here has underpinned much of the in vivo results highlighting the dynamic nature of the plant secretory pathway. Transient transformation of tobacco leaf epidermal cells is a relatively fast technique to assess expression of genes of interest. These cells can be used to generate stable plant lines using a more time-consuming, cell culture technique. Transient expression takes from 2 to 4 days whereas stable lines are generated after approximately 2 to 4 months.

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Citations
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Journal ArticleDOI

Redox-sensitive GFP in Arabidopsis thaliana is a quantitative biosensor for the redox potential of the cellular glutathione redox buffer.

TL;DR: The results with roGFP2 as an artificial GRX target further suggest that redox-triggered changes of biologic processes might be linked directly to the glutathione redox potential via GRX as the mediator.
Journal ArticleDOI

Regulation of Floral Patterning by Flowering Time Genes

TL;DR: The finding of coordinated regulation of SEP3 by flowering time genes reveals a hitherto unknown genetic pathway that prevents premature differentiation of floral meristems and determines the appropriate timing of floral organ patterning.
Journal ArticleDOI

Control of grain size and rice yield by GL2-mediated brassinosteroid responses.

TL;DR: The feasibility of modulating specific brassinosteroid responses to improve plant productivity is demonstrated, and a grain-length-associated QTL, GL2, has the potential to improve grain weight and grain yield.
Journal ArticleDOI

Confocal imaging of glutathione redox potential in living plant cells

TL;DR: Specific interaction of both roGFPs with glutaredoxin in vitro strongly suggests that in situ both variants preferentially act as sensors for the glutathione redox potential, suggesting that roG FP2 is a reliable probe for dynamic redox imaging in planta.
References
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Journal ArticleDOI

Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana

TL;DR: The modified method should facilitate high-throughput transformation of Arabidopsis for efforts such as T-DNA gene tagging, positional cloning, or attempts at targeted gene replacement.
Journal ArticleDOI

Gateway-compatible vectors for plant functional genomics and proteomics.

TL;DR: The utility of pEarleyGate destination vectors for the expression of epitope-tagged proteins that can be affinity captured or localized by immunofluorescence microscopy is demonstrated.
Journal ArticleDOI

Agrobacterium-mediated transformation of Arabidopsis thaliana using the floral dip method

TL;DR: The protocol routinely used in the laboratory for the floral dip method for Arabidopsis transformation is described, which can be routinely obtained and a minimum of several hundred independent transgenic lines generated from just two pots of infiltrated plants within 2–3 months.
Journal ArticleDOI

Visualization of protein interactions in living plant cells using bimolecular fluorescence complementation.

TL;DR: The implementation of a bimolecular fluorescence complementation technique for visualization of protein-protein interactions in plant cells revealed a remarkable signal fluorescence intensity of interacting protein complexes as well as a high reproducibility and technical simplicity of the method in different plant systems.
Journal ArticleDOI

Agrobacterium tumefaciens-mediated transformation of Arabidopsis thaliana root explants by using kanamycin selection.

TL;DR: A transformation procedure for Arabidopsis root explants based on kanamycin selection was established and an Agrobacterium tumor-inducing Ti plasmid carrying a chimeric neomycin phosphotransferase II gene (neo) was introduced, resulting in transformed seed-producing plants obtained with an efficiency between 20% and 80% within 3 months after gene transfer.
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