Conformational, Thermodynamic and Kinetic Studies of RNA-DNA Hydbid Duplexes

本建议的目的是研究如何顺序和 溶液盐浓度影响结构， DNA-RNA杂合寡聚体的热力学和动力学性质。 要解决的问题是：是否存在DNA-RNA杂交双链体 仅作为A型螺旋，或者它们显示依赖于 形状？最近邻序列的作用是什么 杂交双链体的稳定性以及盐浓度如何 影响混合分子的物理性质和行为？ 工作将包括： 1.杂化双链构象的圆二色性研究 层序-盐浓度函数 2.光学熔化研究，以开发自由能的基础集 预测稳定性和构象的参数 1.0 M Na+中的杂交双链体 3.光学熔融法研究盐对 双相形成热力学 4.激光拉曼温度跃变的构造 用分光光度计测量杂化物动力学性质 作为碱基序列和盐的函数的双链体形成 浓度.

The objective of this proposal is to study how sequence and solution salt concentration influence the structural, thermodynamic and kinetic properties of a DNA-RNA hybrid oligomer. Questions to be addressed are: Do DNA-RNA hybrid duplexes exist only as A-form helices or do they display a sequence dependent shape?, What is the role of nearest neighbor sequence determining the stability of a hybrid duplex? and How does salt concentration affect the physical properties and behavior of hybrid molecules? Work will include: 1. Circular dichroism studies of hybrid duplex conformation as a function of sequence and salt concentration 2. Optical melting studies to develop a basis set of free energy parameters for the prediction of stability and conformation of hybrid duplexes in 1.0 M Na+ 3. Optical melting studies of the effects of salt on the thermodynamics of duplex formation 4. The construction of a laser Raman temperature jump spectrophotometer to measure kinetic properties of hybrid duplex formation as a function of base sequence and salt concentration.