論文 - 詳細
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 |