DETECTION OF SPECIFIC RNA BY CONFORMATIONAL CHANGE OF RECOMBINANT PROTEIN
We constructed a novel recombinant protein, which has RNA-binding peptide between ECFP and EYFP fluorescentproteins, for detecting specific RNA-target. We constructed and expressed EYFP- Rev- ECFP recombinant protein in HeLa cells, and confirmed that they could specifically bind to the target - RNA (RRE - RNA or Rev-aptamer) in cell lysate. Subsequently, fluorescent resonance energy transfer (FRET) signal was changed upon RNA binding following the Rev-peptide conformational change. Then we designed RNA-oligonucleotide probes, which can bind to adjacent region of specific RNA-target sequence and reform the Rev-peptide binding site on its hybridrized-complex, for detection of arbitrary sequence RNA-target Finally, specific RNA-target was homogeneously detected in cell lysate.
FRET RNA-peptide interaction RNA-binding protein probe
Eiry Kobatake Tamaki Endoh Masayasu Mie
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology 4259 Nagatsuta-cho, Midori-ku, Yokohama, 226-8503
国际会议
The 4th International Forum on Post-genome Technologies(4IFPT)(第四届国际后基因组生命科学技术学术论坛)
杭州
英文
23-24
2006-09-25（万方平台首次上网日期，不代表论文的发表时间）