基于核酸脱碱基位点的铽离子荧光增强型单核苷酸多态性识别.pdfVIP

2011 年第69 卷 化 学 学 报 Vol. 69, 2011 第18 期, 2137～2142 ACTA CHIMICA SINICA No. 18, 2137～2142 ·研究论文· 基于核酸脱碱基位点的铽离子荧光增强型单核苷酸多态性识别研究 崔清华 邵 勇* 马 坤 刘桂英 吴 飞 许淑娟 (浙江师范大学物理化学研究所 金华 321004) 摘要 基于核酸脱碱基(AP)位点构建了无机配体稀土铽离子(Tb3＋)荧光增强型单核苷酸多态性(SNP)识别方法. 在目标 链靶标碱基对应的探针链上相应位置引入AP 位点, 发现Tb3＋可以选择性地结合在AP 位点, 光激发时发生从DNA 碱 基到结合的Tb3＋的能量转移, 使Tb3＋特征荧光显著增强. 这种荧光增强作用与靶标碱基及AP 位点侧翼碱基类型密切 相关. 当靶标碱基和侧翼碱基为G 时, 荧光最强. 该方法可用于区分肿瘤抑制基因p53 密码子177 位的C/G 碱基变异. 关键词 脱碱基位点; 单核苷酸多态性; 铽离子; 荧光增强; 分子识别 Study on Recognition of Single-nucleotide Polymorphism based on Fluorescence Enhancement of Terbium Ion upon Binding at an Abasic Site in DNA Cui, Qinghua Shao, Yong* Ma, Kun Liu, Guiying Wu, Fei Xu, Shujuan (Institute of Physical Chemistry, Zhejiang Normal University, Jinhua 321004) Abstract Recognition of single-nucleotide polymorphism (SNP) was proposed in this work on the basis of 3＋ fluorescence enhancement of an inorganic ligand, terbium ion (Tb ), upon binding at an abasic (AP) site in DNA. The AP site was deliberately introduced in a probe strand just opposite the base of interest in a target 3＋ strand. Selective binding of Tb at the AP site was found by observations of its characteristic fluorescence enhancement for AP site-containing DNA by comparison with fully matched DNA without the AP site. The enhancement is very dependent on the target base and the bases flanking the AP site. When the target base is 3＋ G, the max