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费氏弧菌Y1菌株的生物发光颜色调制与内源性黄色荧光蛋白水平的改变及其荧光成像

Bioluminescence color modulation of Vibrio fischeri strain Y1 coupled with alterable levels of endogenous yellow fluorescent protein and its fluorescence imaging.

作者信息

Karatani Hajime, Matsumoto Shoji, Miyata Koji, Yoshizawa Susumu, Suhama Yuki, Hirayama Satoshi

机构信息

Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto, Japan.

出版信息

Photochem Photobiol. 2006 Mar-Apr;82(2):587-92. doi: 10.1562/2005-06-29-RA-597.

Abstract

Bioluminescence (BL) (lambda(max) approximately 535 nm) of Vibrio fischeri strain Y1 has been previously characterized in terms of the fluctuation in intracellular levels of yellow fluorescent protein (YFP). In this study fluorescence microscopic analysis has revealed that yellow fluorescence, as well as blue fluorescence attributable to a luciferase intermediate, is localized to the periphery of V. fischeri Y1 cells. This finding indicates that both YFP and the luciferase are present in the vicinity of the cell membrane. By using cyanide to enhance yellow BL, it has been shown that BL modulation is coupled with the fluctuations in the intracellular levels of YFP and the primary emitter. On the basis of the BL characterization, combined with results of a sedimentation experiment, it has been shown that larger cells produce a relatively stronger yellow BL. Two-dimensional gel electrophoresis of cell-protein extracts has shown that the YFP level is more alterable than the luciferase level. It is postulated that the yellow BL modulation takes place in connection with cell growth.

摘要

费氏弧菌Y1菌株的生物发光(BL)(最大波长约535nm)先前已根据黄色荧光蛋白(YFP)细胞内水平的波动进行了表征。在本研究中,荧光显微镜分析表明,黄色荧光以及归因于荧光素酶中间体的蓝色荧光定位于费氏弧菌Y1细胞的周边。这一发现表明YFP和荧光素酶都存在于细胞膜附近。通过使用氰化物增强黄色生物发光,已表明生物发光调节与YFP和主要发射体的细胞内水平波动相关。基于生物发光表征,并结合沉降实验结果,已表明较大的细胞产生相对较强的黄色生物发光。细胞蛋白提取物的二维凝胶电泳表明,YFP水平比荧光素酶水平更易变化。据推测,黄色生物发光调节与细胞生长有关。

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