Scanning of guanine−guanine mismatches in DNA by synthetic ligands
using surface plasmon resonance
Kazuhiko Nakatani, Shinsuke Sando
& Isao Saito
Department of Synthetic Chemistry and Biological Chemistry,
Faculty of Engineering, Kyoto University, CREST, Japan Science and Technology
Corporation (JST), Kyoto 606-8501, Japan
.
Correspondence should be addressed to Kazuhiko Nakatani nakatani@sbchem.kyoto-u.ac.jp G-G mismatchsynthetic ligandsurface plasmon resonanceSNPsmutation detection
Here we have designed and synthesized ligands that specifically bind with
high affinity (Kd = 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.
