通过荧光相关光谱法评估分离的大肠杆菌类核的内部结构和动力学。
Internal structure and dynamics of isolated Escherichia coli nucleoids assessed by fluorescence correlation spectroscopy.
作者信息
Romantsov Tatyana, Fishov Itzhak, Krichevsky Oleg
机构信息
Department of Life Sciences, and Physics Department and Ilse Kats Center for Nanoscience, Ben-Gurion University, Beer-Sheva, Israel.
出版信息
Biophys J. 2007 Apr 15;92(8):2875-84. doi: 10.1529/biophysj.106.095729. Epub 2007 Jan 26.
Abstract
The morphology and dynamics of DNA in a bacterial nucleoid affects the kinetics of such major processes as DNA replication, gene expression. and chromosome segregation. In this work, we have applied fluorescence correlation spectroscopy to assess the structure and internal dynamics of isolated Escherichia coli nucleoids. We show that structural information can be extracted from the amplitude of fluorescence correlation spectroscopy correlation functions of randomly labeled nucleoids. Based on the developed formalism we estimate the characteristic size of nucleoid structural units for native, relaxed, and positively supercoiled nucleoids. The degree of supercoiling was varied using the intercalating agent chloroquine and evaluated from fluorescence microscopy images. The relaxation of superhelicity was accompanied by 15-fold decrease in the length of nucleoid units (from approximately 50 kbp to approximately 3 kbp).
摘要
细菌类核中DNA的形态和动力学影响着DNA复制、基因表达和染色体分离等主要过程的动力学。在这项工作中，我们应用荧光相关光谱法来评估分离出的大肠杆菌类核的结构和内部动力学。我们表明，可以从随机标记类核的荧光相关光谱相关函数的振幅中提取结构信息。基于所建立的形式体系，我们估计了天然、松弛和正超螺旋类核的类核结构单元的特征尺寸。使用嵌入剂氯喹改变超螺旋程度，并从荧光显微镜图像中进行评估。超螺旋松弛伴随着类核单元长度15倍的减少（从约50 kbp降至约3 kbp）。
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