Static and kinetic site-specific protein-DNA photocrosslinking: analysis of bacterial transcription initiation complexes.

Static and kinetic site-specific protein-DNA photocrosslinking: analysis of bacterial transcription initiation complexes.
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DOI:
10.1007/978-1-60327-015-1_25
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发表时间:
2009
期刊:
Methods in molecular biology (Clifton, N.J.)
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静态位点特异性蛋白质-DNA光交联允许鉴定多蛋白质-DNA复合物内的蛋白质-DNA相互作用。动力学位点特异性蛋白质-DNA光交联-涉及快速淬灭流混合和脉冲激光照射-允许阐明多蛋白质-DNA复合物内蛋白质-DNA相互作用形成的途径和动力学。我们提出了详细的协议,应用静态和动态位点特异性蛋白质-DNA光交联细菌转录起始复合物。
Static site-specific protein-DNA photocrosslinking permits identification of protein-DNA interactions within multiprotein-DNA complexes. Kinetic site-specific protein-DNA photocrosslinking--involving rapid-quench-flow mixing and pulsed-laser irradiation--permits elucidation of pathways and kinetics of formation of protein-DNA interactions within multiprotein-DNA complexes. We present detailed protocols for application of static and kinetic site-specific protein-DNA photocrosslinking to bacterial transcription initiation complexes.