DNA DESIGNER CRYSTALS -- 3D TRIANGLE, DODECAMER, 16MER

DNA 设计师水晶 -- 3D 三角形、十二角、16MER

• 批准号: 7957298
• 负责人: JENS J BIRKTOFT
• 金额: $ 1.14万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957298
• 关键词: Behavior; Binding; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Crystallography; DNA; DNA Fingerprinting; Diffusion; Funding; Gel; Grant; Heating; High Pressure Liquid Chromatography; Institution; Laboratories; Length; Light; Methods; Modification; Periodicity; Process; Protocols documentation; Research; Research Personnel; Resolution; Resources; Roentgen Rays; Source; Structure; Suggestion; Synchrotrons; Temperature; United States National Institutes of Health; Work; X ray diffraction analysis; X-Ray Diffraction; design; improved; macromolecule; melting; scaffold; vapor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We have prepared three types of DNA crystals. The first one is called triangle. The space group is R3. Resolution we got is about 5A. The second one is dodecamer DNA crystal. The space group is C2. The resolution we can get for it is about 3A. The third one is 16mer DNA crystal. The space group is P43. The resolution is about 3.1A. Our group has designed a variety of DNA motifs expected to form 3D lattices. Ultimately, the applications of 3D DNA lattices include providing scaffolding that is capable of binding and orienting a variety of species. Crystals of macromolecules and notoriously difficult to grow, and the process of crystallization relies heavily on pure luck. DNA cages could act as a host to these molecules, aligning them in a periodic fashion necessary for X-ray analysis. The 3D designs pursued in our laboratory are predicated on Watson-Crick complementarities. In order to confirm the formation of 3D DNA lattices, we purify the strands corresponding to those designs and self-assemble them into the desired motifs. It will attempt to form crystalline lattices with the DNA motifs. When crystals appear, they will be characterized by X-ray diffraction methods. Derivatized crystals will be prepared, and the structures solved when the resolution is high enough to warrant such studies. A 3D triangle was made in this work involved incorporating DX molecules to replace the double helices. Crystals of this motif were grown using a combination of vapor diffusion and a heating protocol. They ranged from 100 to 200um in length. A two-tile version of the 3D DX triangle has been designed and the AFM evidence supports its formation. So far, we have completed the DLS characterization of the melting behaviors of the aggregates so as to determine proper temperature ranges and preferred lattice periodicities and to establish a relationship between the melting temperatures established by the DLS and the proper temperature at which to attempt crystallization. In our lab we also tried HPLC purification followed by gel purification. The improved resolution on some simple version encourages us to apply HPLC purification on this motif. Also followed Dr. Sherman¿¿¿¿¿"s suggestion, we modified the motif design to make sure the structure is in less strain. Hopefully, during this course, we would have crystals from this new modification.

该副本是使用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这是调查员的机构。 我们准备了三种类型的DNA晶体。 第一个称为三角形。空间组为R3。我们得到的解决方案大约是5A。 第二个是DodeCamer DNA晶体。空间组为C2。我们可以获得的分辨率大约是3A。 第三个是16mer DNA晶体。空间组为p43。分辨率约为3.1a。 我们的小组设计了各种预期形成3D晶格的DNA图案。最终，3D DNA晶格的应用包括提供能够结合和取向各种物种的脚手架。大分子的晶体和众所周知的生长很难生长，结晶的过程在很大程度上取决于纯粹的幸运。 DNA笼可以充当这些分子的宿主，以X射线分析所需的定期方式对齐它们。在我们的实验室中所追求的3D设计是在沃森 - 克里克互补性上预测的。为了确认3D DNA晶格的形成，我们净化了与这些设计相对应的链，并将它们自组装成所需的基序。它将尝试使用DNA图案形成结晶晶格。当晶体出现时，它们将以X射线衍射方法为特征。将准备衍生化的晶体，当分辨率足够高以保证此类研究时，结构已解决。 在这项工作中进行了一个3D三角形，涉及结合DX分子以取代双螺旋。使用蒸气扩散和加热方案的组合生长该基序的晶体。它们的长度从100到200UM不等。设计了3D DX三角形的两英尺版本，AFM证据支持其形成。到目前为止，我们已经完成了聚集体熔化行为的DLS表征，以确定适当的温度范围和首选的晶格周期，并在DLS建立的熔融温度与尝试结晶的适当温度之间建立关系。在我们的实验室中，我们还尝试了HPLC纯化，然后尝试进行凝胶纯化。改进的一些简单版本的分辨率鼓励我们在此主题上应用HPLC纯化。还遵循Sherman博士€€€��``建议，我们修改了主题设计，以确保结构减少压力。希望在本课程中，我们将获得这种新修饰的晶体。