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利用光化学连接检测单个碱基错配

Detection of one base mismatch using photochemical ligation.

作者信息

Mukae Motoko, Tabara Miyuki, Arslan Pelin, Ihara Toshihiro, Jyo Akinori

机构信息

Department of Applied Chemistry and Biochemistry, Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555, Japan.

出版信息

Nucleic Acids Symp Ser (Oxf). 2007(51):287-8. doi: 10.1093/nass/nrm144.

Abstract

We attached anthracene molecules covalently to one end of oligodeoxyribonucleotides to form three pairs of anthracene-ODN conjugates. The sequences of each pair of the conjugates were designed to bind adjacent sequences of the template with the anthracene units directed such that they stacked each other. The conjugates dimerized only in the presence of the template by light irradiation. We found that the ligation yields largely depended on the substituted position of anthracene groups. Sequence selectivity in the dimerization yield was preserved even in the system with highest reactivity. SNP bases could be discriminated rapidly by combining the techniques of MS spectroscopy.

摘要

我们将蒽分子共价连接到寡脱氧核糖核苷酸的一端,以形成三对蒽-寡脱氧核糖核苷酸缀合物。每对缀合物的序列被设计为与模板的相邻序列结合,蒽单元的方向使得它们相互堆叠。通过光照,缀合物仅在模板存在下二聚化。我们发现连接产率在很大程度上取决于蒽基团的取代位置。即使在反应活性最高的体系中,二聚化产率的序列选择性也得以保留。通过结合质谱光谱技术,可以快速区分单核苷酸多态性碱基。

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