Bimolecular Fluorescence Complementation (BiFC) Analysis of Protein-Protein Interaction: How to Calculate Signal-to-Noise Ratio

被引:15
作者
Kodama, Yutaka [1 ]
Hu, Chang-Deng [2 ]
机构
[1] Utsunomiya Univ, Ctr Biosci Res & Educ, Utsunomiya, Tochigi 321, Japan
[2] Purdue Univ, Dept Med Chem & Mol Pharmacol, W Lafayette, IN 47907 USA
来源
LABORATORY METHODS IN CELL BIOLOGY: IMAGING | 2012年 / 113卷
关键词
LIVING CELLS; SIMULTANEOUS VISUALIZATION;
D O I
10.1016/B978-0-12-407239-8.00006-9
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Bimolecular fluorescence complementation (BiFC) is a technique to visualize protein-protein interactions in living cells, and has been widely used in various model organisms. The principle of the BiFC assay is based on the reconstitution of an intact fluorescent protein. The two non-fluorescent fragments are fused to proteins of interest that may interact. If the two proteins interact, the two non-fluorescent fragments are brought together to reconstitute an intact fluorescent protein. The purpose of this protocol is to calculate signal-to-noise (S/N) ratio in the bimolecular fluorescence complementation (BiFC) assay and to provide a semi-quantitative analysis of protein-protein interaction (PPI) in living cells.
引用
收藏
页码:107 / 121
页数:15
相关论文
共 11 条
[1]   Antiparallel leucine zipper-directed protein reassembly: Application to the green fluorescent protein [J].
Ghosh, I ;
Hamilton, AD ;
Regan, L .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2000, 122 (23) :5658-5659
[2]   Simultaneous visualization of multiple protein interactions in living cells using multicolor fluorescence complementation analysis [J].
Hu, CD ;
Kerppola, TK .
NATURE BIOTECHNOLOGY, 2003, 21 (05) :539-545
[3]   Visualization of interactions among bZip and Rel family proteins in living cells using bimolecular fluorescence complementation [J].
Hu, CD ;
Chinenov, Y ;
Kerppola, TK .
MOLECULAR CELL, 2002, 9 (04) :789-798
[4]   An improved mRFP1 adds red to bimolecular fluorescence complementation [J].
Jach, Guido ;
Pesch, Martina ;
Richter, Klaus ;
Frings, Sabine ;
Uhrig, Joachim F. .
NATURE METHODS, 2006, 3 (08) :597-600
[5]   Biomolecular fluorescence complementation (BiFC) analysis as a probe of protein interactions in living cells [J].
Kerppola, Tom K. .
ANNUAL REVIEW OF BIOPHYSICS, 2008, 37 :465-487
[6]  
Kodama Y, 2012, BIOTECHNIQUES, V53, P285, DOI [10.2144/0000113943, 10.2144/000113943]
[7]   An improved bimolecular fluorescence complementation assay with a high signal-to-noise ratio [J].
Kodama, Yutaka ;
Hu, Chang-Deng .
BIOTECHNIQUES, 2010, 49 (05) :793-+
[8]   Simultaneous visualization of two protein complexes in a single plant cell using multicolor fluorescence complementation analysis [J].
Kodama, Yutaka ;
Wada, Masamitsu .
PLANT MOLECULAR BIOLOGY, 2009, 70 (1-2) :211-217
[9]   Fluorescence complementation: an emerging tool for biological research [J].
Shyu, Y. John ;
Hu, Chang-Deng .
TRENDS IN BIOTECHNOLOGY, 2008, 26 (11) :622-630
[10]   Visualization of protein interactions in living Caenorhabditis elegans using bimolecular fluorescence complementation analysis [J].
Shyu, Y. John ;
Hiatt, Susan M. ;
Duren, Holli M. ;
Ellis, Ronald E. ;
Kerppola, Tom K. ;
Hu, Chang-Deng .
NATURE PROTOCOLS, 2008, 3 (04) :588-596