Imaging of Streptomyces coelicolor A3(2) with reduced autofluorescence reveals a novel stage of FtsZ localization.
Imaging of Streptomyces coelicolor A3(2) with reduced autofluorescence reveals a novel stage of FtsZ localization.
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DOI：
10.1371/journal.pone.0004242
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发表时间：
2009
期刊：
影响因子：
3.7
通讯作者：
van Wezel GP
中科院分区：
文献类型：
--
作者：
Willemse J;van Wezel GP
利用荧光显微镜对低丰度蛋白质在时间和空间上的成像通常受到宿主细胞自身荧光的阻碍。链霉菌是研究细菌发育的重要模式系统，在产孢期进行多个同步细胞分裂。为了详细分析这一现象，荧光显微镜，特别是最近发表的新型实时成像技术，需要最佳的信噪比。在这里，我们描述了一种新型衍生品的发展，这种衍生品具有强烈降低的自身荧光，允许以更高的分辨率对荧光标记的蛋白质进行成像。增强的图像细节为细胞分裂蛋白FtsZ提供了新的定位信息，显示了一个新的发育阶段，多个FtsZ病灶聚集在间隔平面上。这表明多个焦点依次产生，最终连接形成完整的Z环。增强的成像特性是对不太丰富的蛋白质进行共聚焦和实时成像以及在链霉菌中使用低强度荧光团的重要一步。
Imaging of low abundance proteins in time and space by fluorescence microscopy is typically hampered by host-cell autofluorescence. Streptomycetes are an important model system for the study of bacterial development, and undergo multiple synchronous cell division during the sporulation stage. To analyse this phenomenon in detail, fluorescence microscopy, and in particular also the recently published novel live imaging techniques, require optimal signal to noise ratios. Here we describe the development of a novel derivative of Streptomyces coelicolor A3(2) with strongly reduced autofluorescence, allowing the imaging of fluorescently labelled proteins at significantly higher resolution. The enhanced image detail provided novel localization information for the cell division protein FtsZ, demonstrating a new developmental stage where multiple FtsZ foci accumulate at the septal plane. This suggests that multiple foci are sequentially produced, ultimately connecting to form the complete Z ring. The enhanced imaging properties are an important step forward for the confocal and live imaging of less abundant proteins and for the use of lower intensity fluorophores in streptomycetes.
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影响因子：
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