RRC ID 45132
著者 Ohashi K, Mizuno K.
タイトル A novel pair of split venus fragments to detect protein-protein interactions by in vitro and in vivo bimolecular fluorescence complementation assays.
ジャーナル Methods Mol Biol
Abstract Protein-protein interactions are critical components of almost every cellular process. The bimolecular fluorescence complementation (BiFC) method has been used to detect protein-protein interactions in both living cells and cell-free systems. The BiFC method is based on the principle that a fluorescent protein is reassembled from its two complementary non-fluorescent fragments when an interaction occurs between two proteins, each one fused to each fragment. In vivo and in vitro BiFC assays, which use a new pair of split Venus fragments composed of VN210 (amino acids 1-210) and VC210 (amino acids 210-238), are useful tools to detect and quantify various protein-protein interactions (including the cofilin-actin and Ras-Raf interactions) with high specificity and low background fluorescence. Moreover, these assays can be applied to screen small-molecule inhibitors of protein-protein interactions.
巻・号 1174
ページ 247-62
公開日 2014-1-1
DOI 10.1007/978-1-4939-0944-5_17
PMID 24947387
MeSH Animals Bacterial Proteins / chemistry Bacterial Proteins / genetics Bacterial Proteins / metabolism* Cell Line Drug Discovery Gene Expression High-Throughput Screening Assays Humans Luminescent Proteins / chemistry Luminescent Proteins / genetics Luminescent Proteins / metabolism* Microscopy, Fluorescence* / methods Plasmids / genetics Protein Binding / drug effects Protein Interaction Mapping* / methods Proteins / chemistry Proteins / genetics Proteins / metabolism* Recombinant Proteins / genetics Recombinant Proteins / isolation & purification Recombinant Proteins / metabolism
引用数 4
WOS 分野 BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
ヒト・動物細胞 HeLa