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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中央赠款提供的资源 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此，在列出的机构中可能会被压制 对于中心，这不一定是调查员的工厂。 我们已经准备了三种类型的DNA晶体。 第一个被称为三角形。 第二个是DNA晶体。 第三个I 16mer DNA晶体是P43。 我们的小组设计了各种DNA图案，形成了3D晶格，能够结合和定向多种物种。分析我们的3D设计是在watson-crick互补性的基础上，以确认3D DNA晶格的形成，我们将与这些设计相对应的链条净化图案。当出现晶体时，将准备X射线衍射方法。 在这项工作中进行了3D三角形，以替换蒸气扩散的离子。支撑骨料的形成，以确定适当的温度范围，范围范围是晶格周期性，并在适当的温度中建立的熔化温度之间的关系。用于在此基序上应用HPLIFIFIEN挥之不去挥之不去挥之不去挥之不去“建议，缪斯女神的主题设计较小。新修改。