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  • 标题:Probing the mechanical stability of bridged DNA-H-NS protein complexes by single-molecule AFM pulling
  • 本地全文:下载
  • 作者:Yan Liang ; Ramon A. van der Valk ; Remus T. Dame
  • 期刊名称:Scientific Reports
  • 电子版ISSN:2045-2322
  • 出版年度:2017
  • 卷号:7
  • 期号:1
  • 页码:15275
  • DOI:10.1038/s41598-017-15477-4
  • 语种:English
  • 出版社:Springer Nature
  • 摘要:Atomic force microscopy (AFM) has proven to be a powerful tool for the study of DNA-protein interactions due to its ability to image single molecules at the nanoscale. However, the use of AFM in force spectroscopy to study DNA-protein interactions has been limited. Here we developed a high throughput, AFM based, pulling assay to measure the strength and kinetics of protein bridging of DNA molecules. As a model system, we investigated the interactions between DNA and the Histone-like Nucleoid-Structuring protein (H-NS). We confirmed that H-NS both changes DNA rigidity and forms bridges between DNA molecules. This straightforward methodology provides a high-throughput approach with single-molecule resolution which is widely applicable to study cross-substrate interactions such as DNA-bridging proteins.
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