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eng
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Author |
Mst Momtaz, Sultana
Department of Molecular and Functional Genomics, Interdisciplinary Center for Science 8 Research, Shimane University, Matsue, Japan
Amit Kumar, Dutta
Department of Molecular and Functional Genomics, Interdisciplinary Center for Science 8 Research, Shimane University, Matsue, Japan
Tanaka, Yuji
Department of Molecular and Functional Genomics, Interdisciplinary Center for Science 8 Research, Shimane University, Matsue, Japan
Mostafa, Aboulela
Department of Molecular and Functional Genomics, Interdisciplinary Center for Science 8 Research, Shimane University, Matsue, Japan
Sugiura, Sayaka
Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
Niwa, Tomoko
Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
Maeo, Kenichiro
Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
Goto-Yamada, Shino
Malopolska Centre of Biotechnology, Jagiellonian University, Krakow, Poland
Kimura, Tetsuya
Department of Life Sciences, Graduate School of Bioresources, Mie University, Tsu, Japan
Ishiguro, Sumie
Department of Applied Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, Japan
Mano, Shoji
Department of Cell Biology, National Institute for Basic Biology, Okazaki, Japan
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Description | Fluorescent proteins are valuable tools in the bioscience field especially in subcellular localization analysis of proteins and expression analysis of genes. Fusion with organelle targeting signal accumulates fluorescent proteins in specific organelles, increases local brightness, and highlights the signal of fluorescent proteins even in tissues emitting a high background of autofluorescence. For these advantages, organelle-targeted fluorescent proteins are preferably used for promoter:reporter assay to define organ-, tissue-, or cell-specific expression pattern of genes in detail. In this study, we have developed a new series of Gateway cloning technology-compatible binary vectors, pGWBs (attR1-attR2 acceptor sites) and R4L1pGWB (attR4-attL1 acceptor sites), carrying organelle-targeted synthetic green fluorescent protein with S65T mutation (sGFP) (ER-, nucleus-, peroxisome-, and mitochondria-targeted sGFP) and organelle-targeted tag red fluorescent protein (TagRFP) (nucleus-, peroxisome-, and mitochondria-targeted TagRFP). These are available for preparation of promoter:reporter constructs by an LR reaction with a promoter entry clone attL1-promoter-attL2 (for pGWBs) or attL4-promoter-attR1 (for R4L1pGWBs), respectively. A transient expression experiment with particle bombardment using cauliflower mosaic virus 35S promoter-driven constructs has confirmed the correct localization of newly developed organelle-targeted TagRFPs by a co-localization analysis with the previously established organelle-targeted sGFPs. More intense and apparent fluorescence signals were detected by the nucleus- and peroxisome-targeted sGFPs than by the normal sGFPs in the promoter assay using transgenic Arabidopsis thaliana. The new pGWBs and R4L1pGWBs developed here are highly efficient and may serve as useful platforms for more accurate observation of GFP and RFP signals in gene expression analyses of plants.
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Subject | promoter assay
organelle-targeted fluorescent protein
binary vector
plant 66 transformation
Gateway cloning.
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Journal Title |
Journal of biotechnology
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Volume | 297
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Start Page | 19
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End Page | 27
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ISSN | 0168-1656
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ISSN(Online) | 1873-4863
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Published Date | 2019-05-19
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DOI | |
PubMed ID | |
Publisher | Amsterdam : Elsevier Science Publishers, c1984-
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NII Type |
Journal Article
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Format |
PDF
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Text Version |
著者版
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Gyoseki ID | e35979
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OAI-PMH Set |
Interdisciplinary Center for Science Research
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