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Subcellular Localization of Interacting Proteins by Bimolecular Fluorescence Complementation in Planta
Year:
2006
Source of publication :
Journal of Molecular Biology
Authors :
Gafni, Yedidya
;
.
Glick, Efrat
;
.
Volume :
362
Co-Authors:
Citovsky, V., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Lee, L.-Y., Department of Biological Sciences, Purdue University, 915 W. State Street, West Lafayette, IN 47907-1392, United States
Vyas, S., Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1048, United States
Glick, E., Institute of Plant Sciences, A.R.O., The Volcani Center, Bet Dagan, 50250, Israel
Chen, M.-H., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Vainstein, A., The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot, 76100, Israel
Gafni, Y., Institute of Plant Sciences, A.R.O., The Volcani Center, Bet Dagan, 50250, Israel
Gelvin, S.B., Department of Biological Sciences, Purdue University, 915 W. State Street, West Lafayette, IN 47907-1392, United States
Tzfira, T., Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1048, United States
Facilitators :
From page:
1120
To page:
1131
(
Total pages:
12
)
Abstract:
Bimolecular fluorescence complementation (BiFC) represents one of the most advanced and powerful tools for studying and visualizing protein-protein interactions in living cells. In this method, putative interacting protein partners are fused to complementary non-fluorescent fragments of an autofluorescent protein, such as the yellow spectral variant of the green fluorescent protein. Interaction of the test proteins may result in reconstruction of fluorescence if the two portions of yellow spectral variant of the green fluorescent protein are brought together in such a way that they can fold properly. BiFC provides an assay for detection of protein-protein interactions, and for the subcellular localization of the interacting protein partners. To facilitate the application of BiFC to plant research, we designed a series of vectors for easy construction of N-terminal and C-terminal fusions of the target protein to the yellow spectral variant of the green fluorescent protein fragments. These vectors carry constitutive expression cassettes with an expanded multi-cloning site. In addition, these vectors facilitate the assembly of BiFC expression cassettes into Agrobacterium multi-gene expression binary plasmids for co-expression of interacting partners and additional autofluorescent proteins that may serve as internal transformation controls and markers of subcellular compartments. We demonstrate the utility of these vectors for the analysis of specific protein-protein interactions in various cellular compartments, including the nucleus, plasmodesmata, and chloroplasts of different plant species and cell types. © 2006 Elsevier Ltd. All rights reserved.
Note:
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carboxy terminal sequence
gene expression
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Plant Breeding
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More details
DOI :
10.1016/j.jmb.2006.08.017
Article number:
0
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
31421
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 01:02
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Scientific Publication
Subcellular Localization of Interacting Proteins by Bimolecular Fluorescence Complementation in Planta
362
Citovsky, V., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Lee, L.-Y., Department of Biological Sciences, Purdue University, 915 W. State Street, West Lafayette, IN 47907-1392, United States
Vyas, S., Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1048, United States
Glick, E., Institute of Plant Sciences, A.R.O., The Volcani Center, Bet Dagan, 50250, Israel
Chen, M.-H., Department of Biochemistry and Cell Biology, State University of New York, Stony Brook, NY 11794-5215, United States
Vainstein, A., The Robert H. Smith Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, Rehovot, 76100, Israel
Gafni, Y., Institute of Plant Sciences, A.R.O., The Volcani Center, Bet Dagan, 50250, Israel
Gelvin, S.B., Department of Biological Sciences, Purdue University, 915 W. State Street, West Lafayette, IN 47907-1392, United States
Tzfira, T., Department of Molecular, Cellular and Developmental Biology, University of Michigan, Ann Arbor, MI 48109-1048, United States
Subcellular Localization of Interacting Proteins by Bimolecular Fluorescence Complementation in Planta
Bimolecular fluorescence complementation (BiFC) represents one of the most advanced and powerful tools for studying and visualizing protein-protein interactions in living cells. In this method, putative interacting protein partners are fused to complementary non-fluorescent fragments of an autofluorescent protein, such as the yellow spectral variant of the green fluorescent protein. Interaction of the test proteins may result in reconstruction of fluorescence if the two portions of yellow spectral variant of the green fluorescent protein are brought together in such a way that they can fold properly. BiFC provides an assay for detection of protein-protein interactions, and for the subcellular localization of the interacting protein partners. To facilitate the application of BiFC to plant research, we designed a series of vectors for easy construction of N-terminal and C-terminal fusions of the target protein to the yellow spectral variant of the green fluorescent protein fragments. These vectors carry constitutive expression cassettes with an expanded multi-cloning site. In addition, these vectors facilitate the assembly of BiFC expression cassettes into Agrobacterium multi-gene expression binary plasmids for co-expression of interacting partners and additional autofluorescent proteins that may serve as internal transformation controls and markers of subcellular compartments. We demonstrate the utility of these vectors for the analysis of specific protein-protein interactions in various cellular compartments, including the nucleus, plasmodesmata, and chloroplasts of different plant species and cell types. © 2006 Elsevier Ltd. All rights reserved.
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