SOLID STATE NMR OF DNA DYNAMICS AND STRUCTURE AND INSTRUMENT DESIGN

我的研究小组将参与四个主要项目。第一 项目是为一个非常高的磁场设计和建造一个控制台 核磁共振波谱仪。一种能在高分辨率下工作的核磁共振操作台的设计 质子频率从750兆赫不等。到1000兆赫。目前有资金 由能源部提出，最终目标是建造1000兆赫。核磁共振 在华盛顿州里奇兰的巴特尔太平洋西北实验室。在.期间 明年，高速采集器的设计和原型版本 系统和PULSE编程器将完成、安装和测试 在旧的NIH计划项目500 MHz上。分光计。微缩VAX 控制该控制台操作的计算机将被替换为 用于控制控制台功能的SGI工作站和软件 也将编写和测试Silicon Graphics工作站。在.期间 在接下来的几年里，我们将使用NIH的资金组装一架750 MHz的飞机。核磁共振 将整合所有这些设计增强功能的控制台。 作为第二个项目，我们将继续研究内部动力学 用~1H、~(13)C和~(15)N固体核磁共振法研究DNA低聚体 光谱学是我们的主要工具。在DT基础上的各种地点 含TnAn的DNA低聚物中的da将被13C和15N富集化 为了区分运动轴的方向， 特定CSA张量的不对称性和各向异性的平均值。 最终，固态核磁共振技术将被扩展到对 DNA三联体和Holliday连接的动力学。 作为第三个项目，我们提出了固体核磁共振的第一个应用 结构技术，如Redor，戏剧和Sedor(RFDR)到 含AnTn的DNA低聚体中核间距的测量。 我们对探索结构特征的本质感兴趣，这些结构特征 导致B‘螺旋的形成，特别是我们打算使用 固体核磁共振作为碱在碱中相互取向的证明 碱基对和相邻碱基之间。 我们的第四个项目涉及合成标记的DNA寡聚体。 在100-200年间合成了选择性的氢化DNA低聚物 毫克量，我们已经成功地合成了150 [d(CGAATTCGCG)]2毫克选择性地标记在C4、N1和 T7和T8的N3位。在不久的将来，我们打算合成DT 和Da，其糖环标记为C1‘、C2’、C3‘和C5’。

My research group will be involved in four major projects. The first project is the design and construction of a console for a very high field NMR spectrometer. The design of an NMR console capable of operating at proton frequencies ranging from 750 MHz. to 1000 MHz. is currently funded by the DOE, with the ultimate objective of construction a 1000 MHz. NMR at the Battelle pacific Northwest Laboratory in Richland, WA. During the next year, designs and prototype versions of the high speed acquisition system and the pulse programmer will be completed, installed and tested on the old NIH Program Project 500 MHz. spectrometer. The microVax computer which controls that console's operation will be replaced with a SGI workstation and software for controlling console function via a Silicon Graphics workstation will also be written and tested. During the next several years we will use NIH funding to assemble a 750MHz. NMR console which will incorporate all of these design enhancements. As a second project, we will continue our study of the internal dynamics of DNA oligomers using deuterium (2H), 13C, and 15N solid state NMR spectroscopy as our principle tools. Various sites on the bases of dT and dA in TnAn-containing DNA oligomers will be enriched with 13C and 15N in an attempt to distinguish the orientation of motional axes for the averaging of the asymmetry and the anisotropy of specific CSA tensors. Ultimately, solid state NMR techniques will be extended to the study of dynamics in DNA triplexes and Holliday junctions. As a third project, we proposed the first application of solid state NMR structural techniques like REDOR, DRAMA, and SEDOR(RFDR) to the measurement of internuclear distances in AnTn-containing DNA oligomers. We are interested in exploring the nature of structural features which result in the formation of a B'helix, and specifically we intend to use solid state NMR as a prove of the mutual orientation of bases within base pairs and between adjacent bases. Our fourth project involves the synthesis of labeled DNA oligomers. Selectively deuterated DNA oligomers have been synthesized in 100-200 milligram quantities, and we have succeeded in synthesizing 150 milligrams of [d(CGCGAATTCGCG)]2 selectively labeled at the C4, N1, and N3 positions of T7 and T8. In the near future we intend to synthesize dT and dA with the sugar rings labeled at C1', C2', C3', and C5'.