Combining Unique Multiplex Gateway Cloning and Bimolecular Fluorescence Complementation (BiFC) for High-Throughput Screening of Protein–Protein Interactions
نویسندگان
چکیده
منابع مشابه
Detection of protein interactions in plant using a gateway compatible bimolecular fluorescence complementation (BiFC) system.
We have developed a BiFC technique to test the interaction between two proteins in vivo. This is accomplished by splitting a yellow fluorescent protein (YFP) into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest. These constructs can then be co-transformed into Nicotiana benthamiana via Agrobacterium mediated transformation, allowing the transit expression o...
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Yuji Tanaka1, Tetsuya Kimura2, Kazumi Hikino3, Shino Goto3,4, Mikio Nishimura3,4, Shoji Mano3,4 and Tsuyoshi Nakagawa1 1Department of Molecular and Functional Genomics, Center for Integrated Research in Science, Shimane University, 2Department of Sustainable Resource Science, Graduate School of Bioresources, Mie University, 3Department of Cell Biology, National Institute for Basic Biology, 4Dep...
متن کاملMONITORING iN ViVO PROTEIN-PROTEIN INTERACTIONS BY COUPLING BIMOLECULAR FLUORESCENCE COMPLEMENTATION (BIFC) AND FLOW CYTOMETRY
We present a high-throughput approach to study weak protein-protein interactions by coupling bimolecular fluorescent complementation (BiFC) to flow cytometry (FC). Bimolecular Fluorescence Complementation (BIFC) is a method to detect proteinprotein interactions based on the formation of a fluorescent complex by fragments of the yellow fluorescent protein (NYFP and CYFP) brought together by the ...
متن کاملBimolecular fluorescence complementation (BiFC) analysis of protein interactions in live cells.
Bimolecular fluorescence complementation (BiFC) analysis enables direct visualization of protein interactions and modifications in living cells. It is based on the facilitated association of two nonfluorescent fragments of a fluorescent protein fused to putative interaction partners. The intrinsic fluorescence of the active complex enables detection of protein interactions with high sensitivity...
متن کاملBimolecular fluorescence complementation (BiFC) analysis as a probe of protein interactions in living cells.
Protein interactions are a fundamental mechanism for the generation of biological regulatory specificity. The study of protein interactions in living cells is of particular significance because the interactions that occur in a particular cell depend on the full complement of proteins present in the cell and the external stimuli that influence the cell. Bimolecular fluorescence complementation (...
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ژورنال
عنوان ژورنال: Journal of Biomolecular Screening
سال: 2016
ISSN: 1087-0571,1552-454X
DOI: 10.1177/1087057116659438