Visualization of Phosphoinositides via the Development of the Transient Expression System of a Cyan Fluorescent Protein in the Red Alga Porphyra yezoensis

被引:0
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作者
Koji Mikami
Toshiki Uji
Lin Li
Megumu Takahashi
Hajime Yasui
Naotsune Saga
机构
[1] Hokkaido University,Faculty of Fisheries Sciences
[2] Hokkaido University,Graduate School of Fisheries Sciences
来源
Marine Biotechnology | 2009年 / 11卷
关键词
Pleckstrin homology domain; Phosphoinositide; Cyan fluorescent protein; Transient gene expression;
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中图分类号
学科分类号
摘要
Phosphoinositides (PIs) play important roles in signal transduction pathways and the regulation of cytoskeleton and membrane functions in eukaryotes. Subcellular localization of individual PI derivative is successfully visualized in yeast, animal, and green plant cells using PI derivative-specific pleckstrin homology (PH) domains fused with a variety of fluorescent proteins; however, expression of fluorescent proteins has not yet been reported in any red algal cells. In the present study, we developed the system to visualize these PIs using human PH domains fused with a humanized cyan fluorescent protein (AmCFP) in the red alga Porphyra yezoensis. Plasma membrane localization of AmCFP fused with the PH domain from phospholipase Cδ1 and Akt1, but not Bruton’s tyrosine kinase, was observed in cell wall-free monospores, demonstrating the presence of phosphatidylinositol-4,5-bisphosphate and phosphatidylinositol-3,4-bisphosphate in P. yezoensis cells. This is the first report of the successful expression of fluorescent protein and the monitoring of PI derivatives in red algal cells. Our system, based on transient expression of AmCFP, could be applicable for the analysis of subcellular localization of other proteins in P. yezoensis and other red algal cells.
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页码:563 / 569
页数:6
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