Abstract
We have developed an optimized protocol for isolating protoplasts from chlorenchyma cells of the single-cell C4 species Bienertia sinuspersici. The isolated protoplasts maintained the integrity of the unique single-cell C4 intracellular compartmentation of organelles as observed in chlorenchyma cells after cell wall digestion. Approximately over 80% of isolated protoplasts expressed the fusion reporter gene following the polyethylene glycol-mediated transfection procedures. Overall, fluorescent protein fusion tagged with various intraorganellular sorting signals validated the potential use of the transient gene expression system in subcellular localization and organelle dynamics studies.
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Acknowledgments
This research was supported by Discovery Grants from the Natural Sciences and Engineering Research Council of Canada (NSERC) RGPIN-2017-04416 to SDXC.
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Lung, SC., Yogadasan, N., Yanagisawa, M., Chuong, S.D.X. (2022). Protoplast Isolation and Transfection in the Single-Cell C4 Species Bienertia sinuspersici . In: Wang, K., Zhang, F. (eds) Protoplast Technology. Methods in Molecular Biology, vol 2464. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2164-6_2
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DOI: https://doi.org/10.1007/978-1-0716-2164-6_2
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-2164-6
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