Visualization of oxidative phosphorylation - reactive oxygen species and its application to self-organization of mitochondrion

2015 Fiscal Year Final Research Report

• Project/Area Number: 25440068
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Biophysics
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: Karatani Hajime 京都工芸繊維大学, 分子化学系, 教授 (10169659)
• Co-Investigator(Renkei-kenkyūsha): OYAMA Hiroshi 摂南大学, 理工学部, 教授 (50221700)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 生物発光 / 発光細菌 / 大腸菌 / ミトコンドリア / 酸化的リン酸化 / 活性酸素種 / ルシフェラーゼ / 蛍光タンパク質

Outline of Final Research Achievements

Results obtained are summarized as follows: (i) From the fluorescent characterization of the recombinant MT-Y1-Yellow and MT-Y1-Blue originating from luminous bacterium, it was found that the mitochondrial signal sequence (MT) does not affect their fluorescence properties; (ii) the expression systems for bacterial luciferase coding gene cassette and firefly luciferase coding gene were improved to yield these luciferases efficiently and these luciferase coding genes were fused; (iii) hydrogen peroxide sensitive genes, both of which are leading to the production of either bioluminescence or fluorescence, have been developed and applied for visualization of hydrogen peroxide in living Escherichia coli; (iv) Based on the further investigation into the bioluminescence ring pattern appearing in the colony of the bioluminescent E. coli, it was found that there is a possibility that the respiratory activity in living cells can be characterized on the basis of bioluminescence.

Free Research Field

生物分子科学