2004 Fiscal Year Final Research Report Summary
Study on molecular mechanism for modulation of bioluminescence from luminous bacteria and molecular function of its fluorescent protein.
| Project/Area Number |
15510173
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Living organism molecular science
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
KARATANI Hajime Kyoto Institute of Technology, Department of Polymer Science and Engineering, Associate Professor, 繊維学部, 助教授 (10169659)
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| Co-Investigator(Kenkyū-buntansha) |
WADA Minoru University of Tokyo, Ocean Research Institute, Research Associate, 海洋研究所, 助手 (70292860)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Luminous bacterium / Bioluminescence modulation / Luciferase / Yellow fluorescent protein / Blue fluorescent protein / Gene cloning / Fluorescence imaging / Cell division cycle |
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| Research Abstract |
It has been revealed that the modulation of bioluminescence(BL) arising from Vibrio fischeri Y1 is coupled with the respiratory protein complex in cell membrane. The BL modulation-respiration coupling seems to be mainly attributed to the electron exchange interaction between endogenous yellow fluorescent protein (YFP) and the respiratory electron flow. Furthermore the BL modulation due to the correlation with the respiration has been demonstrated in a single cell by making use of both the fluorescence microscopy and BL spectroscopy.
Based on electrophoretic analysis, the fluctuation in the intracellular level of endogenous blue fluorescent protein (Y1-BFP) as well as YFP to that of luciferase is a dominant factor to control the magnitude of yellow BL. Fluorescence imaging also has shown that V.fischeri Y1 cell carries both YFP and Y1-BFP simultaneously. The analysis of BL behavior and fluorescence images of the cells fractionated based on a single cell density has made clear that the cells with the smallest density, which seems to be in the phase just before cytokinesis, emit the most intense yellow BL. From these results, it has been postulated that the BL modulation is synchronized with the cell division cycle (cdc).
The genes encoding YFP and Y1-BFP have been cloned completely. The genes for YFP and Y1-BFP consist of 572 bp and 600 bp, respectively. It has also been shown that the deduced amino acid sequence of YFP differs from that of Y1-BFP. The expressions of the two genes have also been separately accomplished in Escherichia coli cell. Assuming that the development of the BL modulation is associated with the fluctuation in the intracellular level of luciferase and the two fluorescent proteins, there seems to be a time lag in the expression of those genes during cdc to be responsible for the development of the BL modulation.
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