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

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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Figure 1: Expression of fluorescent fusion proteins in tobacco epidermal cells.
Figure 2
Figure 3: N. tabacum leaf infiltration.
Figure 4

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Acknowledgements

We are grateful to members of C. Hawes' and I. Moore's laboratories for fruitful discussions and members of the laboratories who have helped refine these techniques over the years. The work reported here was funded by various BBSRC grants.

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Correspondence to Imogen A Sparkes.

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Sparkes, I., Runions, J., Kearns, A. et al. Rapid, transient expression of fluorescent fusion proteins in tobacco plants and generation of stably transformed plants. Nat Protoc 1, 2019–2025 (2006). https://doi.org/10.1038/nprot.2006.286

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