Leiden Institute of Chemistry, Leiden University, Leiden, Netherlands.
National Institute of Molecular Biology and Biotechnology, University of the Philippines, Quezon City, Philippines.
Methods Mol Biol. 2024;2819:279-295. doi: 10.1007/978-1-0716-3930-6_14.
Atomic force microscopy is a high-resolution imaging technique useful for observing the structures of biomolecular complexes. This approach provides a straightforward method to characterize the binding behavior of different chromatin architectural proteins and to analyze the increasingly complex structural units assembled on the DNA. The protocol describes the preparation, AFM imaging, and structural analysis of chromatin that is reconstituted in vitro using purified proteins and DNA. Here, we describe the successful application of the method on the chromatin architectural proteins of the archaeon Sulfolobus solfataricus.
原子力显微镜是一种高分辨率成像技术,可用于观察生物分子复合物的结构。这种方法提供了一种直接的方法来表征不同染色质结构蛋白的结合行为,并分析在 DNA 上组装的越来越复杂的结构单元。该方案描述了使用纯化蛋白和 DNA 在体外重建的染色质的制备、AFM 成像和结构分析。在这里,我们成功地将该方法应用于古菌 Sulfolobus solfataricus 的染色质结构蛋白。