DNA ELECTION TRANSFER AT HYDROGEL MODIFIED ELECTRODES

水凝胶修饰电极上的 DNA 电子转移

基本信息

批准号：
6329600
负责人：
VERONIKA Ann SZALAI
金额：
$ 2.99万
依托单位：
UNIV OF NORTH CAROLINA CHAPEL HILL
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-12-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6329600
关键词：
DNA analytical chemistry bioengineering /biomedical engineering biomaterials biosensor device carrageenan electrochemistry electrodes electron transport gel guanine nucleic acid probes nucleic acid quantitation /detection oxidation ruthenium technology /technique development

项目摘要

As an indicator of diseases like cancer, DNA is a natural target for development of biosensors. Of particular biological and medical interest are sequences derived from infectious organisms or oncogenic cells. Dr. H. Holden Thorp's laboratory has demonstrated that femtomolar quantities of DNA sequences important in cancer research can be electrochemically detected through oxidation of guanine in DNA by the soluble inorganic complex, tris (2.2'-bipyridine)ruthenium(II) - [Ru(bpy)3]2+. The long-term objective of the research proposed here is to develop an electrochemical DNA-detection assay using a carrageenan hydrogel matrix to confine Ru(bpy)32+ and DNA so that neither species can migrate away from the electrode surface. Specific aim (i) is to electrochemically detect guanine oxidation by Ru(bpy)32+ in a carrageenan hydrogel. This specific aim requires incorporation of guanine into a carrageenan hydrogel containing Ru(bpy)32+, determination of the optimal electrode system, and investigation of the kinetics of guanine oxidation in carrageenan hydrogels. Specific aim (ii) is to develop an electrochemical DNA-detection system using a carrageenan hydrogel and Ru(bpy)32+. This specific aim requires attachment of oligonucleotide probes to the ruthenium-doped carrageenan hydrogel followed by quantification of guanine oxidation. The method for probe/target incorporation into the hydrogel will rely on covalent attachment of single-stranded probes or the use of hybridized biotinylated-probe/target complexes with avidin-labeled carrageenan.

作为癌症等疾病的指标，DNA 是生物传感器开发的天然目标。特别具有生物学和医学意义的是源自传染性生物体或致癌细胞的序列。 H. Holden Thorp 博士的实验室证明，通过可溶性无机复合物三 (2.2'-联吡啶)钌 (II) - [Ru( bpy)3]2+。这里提出的研究的长期目标是开发一种电化学 DNA 检测方法，使用角叉菜胶水凝胶基质来限制 Ru(bpy)32+ 和 DNA，以便这两种物质都不能迁移离开电极表面。具体目标 (i) 是电化学检测角叉菜胶水凝胶中 Ru(bpy)32+ 对鸟嘌呤的氧化作用。这一具体目标需要将鸟嘌呤掺入含有 Ru(bpy)32+ 的卡拉胶水凝胶中，确定最佳电极系统，并研究卡拉胶水凝胶中鸟嘌呤氧化的动力学。具体目标 (ii) 是开发一种使用角叉菜胶水凝胶和 Ru(bpy)32+ 的电化学 DNA 检测系统。这一具体目标需要将寡核苷酸探针附着到掺钌角叉菜胶水凝胶上，然后对鸟嘌呤氧化进行定量。将探针/靶标掺入水凝胶的方法将依赖于单链探针的共价连接或使用杂交的生物素化探针/靶标复合物与亲和素标记的角叉菜胶。