RT Journal
A1 Doi, Nobuhide
A1 Takashima, Hideaki
A1 Kinjo, Masataka
A1 Sakata, Kyoko
A1 Kawahashi, Yuko
A1 Oishi, Yuko
A1 Oyama, Rieko
A1 Miyamoto-Sato, Etsuko
A1 Sawasaki, Tatsuya
A1 Endo, Yaeta
A1 Yanagawa, Hiroshi
T1 Novel Fluorescence Labeling and High-Throughput Assay Technologies for In Vitro Analysis of Protein Interactions
JF Genome Research 
JO Genome Research 
YR 2002 
FD March 01 
VO 12 
IS 3 
SP 487 
OP 492 
DO 10.1101/gr.218802 
UL http://genome.cshlp.org/content/12/3/487.abstract 
AB We developed and tested a simple method for fluorescence labeling and interaction analysis of proteins based on a highly efficient in vitro translation system combined with high-throughput technologies such as microarrays and fluorescence cross-correlation spectroscopy (FCCS). By use of puromycin analogs linked to various fluorophores through a deoxycytidylic acid linker, a single fluorophore can be efficiently incorporated into a protein at the carboxyl terminus during in vitro translation. We confirmed that the resulting fluorescently labeled proteins are useful for probing protein–protein and protein–DNA interactions by means of pulldown assay, DNA microarrays, and FCCS in model experiments. These fluorescence assay systems can be easily extended to highly parallel analysis of protein interactions in studies of functional genomics.[Online supplementary material available at http://www.genome.org.]