Scanning of guanine-guanine mismatches in DNA by synthetic ligands using surface plasmon resonance

被引:202
作者
Nakatani, K [1 ]
Sando, S [1 ]
Saito, I [1 ]
机构
[1] Kyoto Univ, Japan Sci & Technol Corp, CREST, Fac Engn,Dept Synth Chem & Biol Chem, Kyoto 6068501, Japan
关键词
G-G mismatch; synthetic ligand; surface plasmon resonance; SNPs; mutation detection;
D O I
10.1038/83505
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Here we have designed and synthesized ligands that specifically bind with high affinity (K-d = 53 nM) to the guanine (G)-guanine mismatch, one of four types of single-nucleotide polymorphism (SNP). Detection of the G-G mismatch was performed by a surface plasmon resonance (SPR) assay using a sensor chip carrying the G-G specific ligand on its surface. The accuracy of the G-G mismatch detection by the SPR sensor was demonstrated by a marked SPR response obtained only for the DNA containing the G-G mismatch. DNAs containing G-A and G-T mismatches, as well as a fully matched duplex, produced only a weak response. Furthermore, this assay was found applicable for the detection of SNP existing in PCR amplification products of a 652-nucleotide sequence of the HSP70-2 gene.
引用
收藏
页码:51 / 55
页数:5
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