Cyclic ferrocenyl intercalator as a novel probe for gene polymorphism analysis

环状二茂铁嵌入剂作为基因多态性分析的新型探针

• 批准号: 13680672
• 负责人: TAKENAKA Shigeori
• 金额: $ 2.24万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13680672/
• 关键词: human genome; SNP; naphthalene diimide; ferrocene; intercalator; cyclic intercalator; electrochemical measurement; molecular probe; アントラキノン; 大環状インターカレータ; 一塩基多型(SNP); 解析プローブ; ハイブリダイゼーション; ヘテロ二重らせん; 融解温度; ナフタレンジイミド; 遺伝子多型; ミスマッチ塩基; X線構造解析

The research on the human genome leads to single nucleotide polymorphism (SNP) analysis eventually at the individual level. The goal of this project was, hence, to develop a cyclic intercalator, CyNDIFc, composed of naphthalene diimide and ferrocene parts as an electrochemical probe for SNP analysis. The integrity of the structure of CyNDIFc newly synthesized was confirmed by X-ray analysis. The effect of CyNDIFc on the mismatched DNA region was evaluated by Tm measurements of a DNA duplex in its presence. The Tm value in the presence of CyNDIFc increased when a mismatched base was introduced in the DNA duplex. Electrochemical measurement was performed by using a DNA-immobilized electrode in order to detect the mismatched base on the DNA duplex. The current from CyNDIFc increased as mismatched base regions increased. These results showed that CyNDIFc is a promising candidate as a new molecular probe for SNP analysis. Further optimization of the CyNDIFc structure will lead to molecular probes with even higher selectivity and sensitivity.

对人类基因组的研究导致单核苷酸多态性（SNP）分析最终在个体水平上进行。因此，本项目的目标是开发一种由萘二酰亚胺和二茂铁部分组成的环状嵌入剂CyNDIFc，作为SNP分析的电化学探针。通过X射线分析证实了新合成的CyNDIFc结构的完整性。CyNDIFc对错配DNA区域的影响通过在其存在下DNA双链体的Tm测量来评价。当在DNA双链体中引入错配碱基时，CyNDIFc存在下的Tm值增加。为了检测DNA双链体上的错配碱基，使用DNA固定化电极进行电化学测量。来自CyNDIFc的电流随着错配碱基区的增加而增加。这些结果表明，CyNDIFc是一个有前途的候选人作为一个新的分子探针的SNP分析。CyNDIFc结构的进一步优化将导致具有更高选择性和灵敏度的分子探针。

项目成果

H. Miyahara, et al.: "Electrochemical analysis of single nucleotide polymorphisms of p53 gene"Talanta. 56. 829-835 (2002)

H.Miyahara等人：“p53基因单核苷酸多态性的电化学分析”Talanta。

K.Yamashita et al.: "Visualization of DNA microarrays by scanning electrochemical microscopy (SECM)"Analyst. 126. 1201-1202 (2001)

K.Yamashita 等人：“通过扫描电化学显微镜 (SECM) 实现 DNA 微阵列的可视化”分析师。

S.Takenaka: "Small Molecule DNA and RNA Binders : From Synthesis to Nucleic Acid"Wiley-VCH. 23 (2003)

S.Takenaka：“小分子 DNA 和 RNA 结合剂：从合成到核酸”Wiley-VCH。

T. Nojima, et al.: ""Direct detection of single nucleotide polymorphism (SNP) with genomic DNA by the ferrocenylnaphthalene diimide-based electrochemical hybridization assay (FND-EHA)"Analytical Sciences. 19. 79-83 (2003)

T. Nojima 等人：“通过基于二茂铁基萘二酰亚胺的电化学杂交测定 (FND-EHA) 用基因组 DNA 直接检测单核苷酸多态性 (SNP)”Analytical Sciences. 19. 79-83 (2003)

S. Takenaka, et al.: "Base mutation analysis by a ferrocenyl naphthalene diimide drivative"Nucleosides, Nucleotides & Nucleic Acids. 20. 1429-1432 (2001)

S. Takenaka 等人：“二茂铁基萘二酰亚胺衍生物的碱基突变分析”核苷、核苷酸