NUCLEASE ACTIVITY OF 1, 10-PHENANTHROLINE-COPPER ION

1，10-二氮杂菲-铜(OP-Cu)镍核酸酶活性的研究 通过对核糖部分的氧化攻击。自由配位络合物是 对研究核酸结构/功能关系很有用。 连接到载体配体上的OP-Cu允许合成具有 精心设计的细节。 将开发基因组图谱的目标核酸酶活性， 涉及a)细菌抑制物的基因工程衍生物 CRO蛋白；以及b)使用经以下修饰的RNA形成R环 1，10-二氮杂菲。这两种方法都将能够在任何选择的情况下被删除 标记基因序列。基因特异性抑制物的合成 转录将通过将OP-铜连接到寡核苷酸a)实现。 与转录起始点的单链区域互补； 或b)能够与多嘌呤/多嘧啶形成形式的三股螺旋 真核基因两侧的区域。 核糖核酸酶的化学机制和二级结构特异性 将测定OP-Cu的活性。以及光激活的铀酰 醋酸酯作为RNA-蛋白质足迹试剂的潜力 本课程将探讨互动。其间形成三股螺旋 化学衍生化的双链RNA和脱氧寡核苷酸 将对OP-Cu进行研究。通过分析目标的断裂模式 RNA，三螺旋形成的严密性、极性和稳定性将 要下定决心。这种交互提供了一种新的设计方法 反义试剂在RNA水平上阻断基因表达。

The nuclease activity of 1,10-phenanthroline-Cu (OP-Cu) nicks RNA and DNA by oxidative attack on the ribose moiety. The free coordination complex is useful for investigating nucleic acid structure/function relationships. Linked to carrier ligands, OP-Cu allows synthesis of nucleases with designed specificities. Targeted nuclease activities for genomic mapping will be developed which involve a) genetically engineered derivatives of the bacterial repressor cro protein; and b) R-loop formation using RNAs modified with 1,10-phenanthroline. Both methods will be capable of scission at any chosen marker gene sequence. The synthesis of gene specific inhibitors of transcription will be achieved by linking OP-Cu to oligonucleotides a) complementary to single stranded regions of transcription initiation sites; or b) capable of formal, triple helices with polypurine/polypyrimidine tracts which flank eucaryotic genes. The chemical mechanism and secondary structure specificity of the RNase activity of OP-Cu will be determined. Along with light-activated uranyl acetate, its potential as a footprinting reagent for RNA-protein interactions will be explored. Triple helix formation between double-stranded RNA and deoxyoligonucleotides chemically derivatized with OP-Cu will be investigated. By analyzing the scission pattern of the target RNA, the stringency, polarity, and stability of triple helix formation will be determined. This interaction provides a new approach for the design of antisense reagents to block gene expression at the level of RNA.