Study on molecular mechanism for modulation of bioluminescence from luminous bacteria and molecular function of its fluorescent protein.

发光细菌生物发光调节的分子机制及其荧光蛋白的分子功能研究。

• 批准号: 15510173
• 负责人: KARATANI Hajime
• 金额: $ 2.05万
• 依托单位: Kyoto Institute of Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15510173/
• 关键词: Luminous bacterium; Bioluminescence modulation; Luciferase; Yellow fluorescent protein; Blue fluorescent protein; Gene cloning; Fluorescence imaging; Cell division cycle; 蛍光タンパク質; 生物発光; 呼吸; 呼吸鎖電子伝達系; Luminous bacterium; Bioluminescence modulation; Luciferase; Yellow fluorescent protein; Blue fluorescent protein; Gene cloning; Fluorescence imaging; Cell division cycle; 蛍光タンパク質; 生物発光; 呼吸; 呼吸鎖電子伝達系

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.

据揭示，由Fischeri Y1引起的生物发光（BL）的调节与细胞膜中的呼吸蛋白复合物耦合。 BL调节示波耦合似乎主要归因于内源性黄色荧光蛋白（YFP）和呼吸电子流之间的电子交换相互作用。 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.荧光成像还表明，V.Fischeri Y1细胞同时携带YFP和Y1-BFP。基于单个细胞密度分离的细胞的BL行为和荧光图像的分析表明，最小密度的细胞似乎在细胞因子之前就处于相位，这会发出最强烈的黄色BL。从这些结果中可以推测，BL调制与细胞分裂周期（CDC）同步。编码YFP和Y1-BFP的基因已完全克隆。 YFP和Y1-BFP的基因分别由572 bp和600 bp组成。还已经表明，推论的YFP的氨基酸序列与Y1-BFP的氨基酸序列不同。这两个基因的表达也已在大肠杆菌细胞中分别完成。假设BL调制的发展与荧光素酶和两种荧光蛋白的细胞内水平的波动有关，则CDC期间这些基因的表达似乎存在时间滞后，以负责BL调节的发展。