Development of bioluminescent probe for small compound based on BRET

Research Project

Project/Area Number	25350983
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Multi-year Fund
Section	一般
Research Field	Chemical biology
Research Institution	National Institute of Advanced Industrial Science and Technology
Principal Investigator	WU CHUN 国立研究開発法人産業技術総合研究所, バイオメディカル研究部門, 主任研究員 (90415646)
Project Period (FY)	2013-04-01 – 2016-03-31
Project Status	Completed (Fiscal Year 2015)
Budget Amount *help	¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000) Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000) Fiscal Year 2014: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000) Fiscal Year 2013: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Keywords	発光プローブ / ペプチド / ファージ / 生物発光共エネルギー移動 / ルシフェラーゼ / 蛍光タンパク質 / 生物発光共鳴エネルギー移動 / アデノシン三リン酸 / スクリーニング / アプタマー / スクリーニングこ
Outline of Final Research Achievements	In this study, we tried to develop an optical probe for imaging small molecule by using bioluminescence resonance energy transfer (BRET) method and phage display technique. First, two peptides which have affinity for S-adenosyl-homocysteine were selected from phage library. Then the linker peptide between the selected peptides were optimized.The DNA coding the selected peptide was inserted to the DNA construct containing gaussia luciferase and green fluorescent protein genes. Finally the detection of SAH was examined by monitoring of the BRET signal of the fusion protein.