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	Single-nucleotide polymorphisms detection by using abasic site-containing dna duplexes
	Zhao CX(赵春霞) ; Nishizawa ; S. ; Teramae ; N. ; Xu GW(许国旺)
	2007-06-04
会议名称	dalian international symposia and exhibition on chromatography
会议日期	2007-6-4
会议地点	中国
页码	76/2
通讯作者	许国旺
中文摘要	detection of single-nucleotide polymorphisms (snps) in human genome has become increasingly important in the fields of genetics, molecular diagnostics and cancer research. various detection methods for snps, including high-density arrays, primer extension methods, real time pcr, invader assay, and direct sequencing, have been developed and used in basic research, clinical diagnosis of genetic disorders and certain cancers. however, these methods are rather labor-intensive and time-consuming, so the demand still remains for a rapid, specific and cost-effective method for the routine detection of snps. here we present potential use of a hydrogen bond-forming ligand, n-amidino-3,5- diamino-6-chloropyrazinecarboxamide hydrochloride (amiloride), for the fluorescence detection of the thymine (t)-related snps. in contrast to current methods for snps typing, our method is characterized by construction of the abasic site (ap site) in dna duplexes, which allows small ligands to bind to target nucleotides with fluorescence signaling: an ap site-containing probe dna is hybridized with a target dna so as to place the ap site toward a target nucleotide, by which hydrophobic microenvironments are provided for ligands to recognize target nucleotides. in this study, a propyl residue (spacer c3) which lacks a nucleobase moiety is utilized for the design of an ap site and is incorporated with 11-mer odns (5’-tccagxgcaac-3’, x = spacer c3). from the examination of the binding behaviors of amiloride, we find that amiloride selectively binds to t with high affinity (ka = 6.7 × 106 m-1), and this is accompanied by remarkable quenching of its fluorescence. applications of amiloride for the detection of t-related snps of human genes were also demonstrated.
会议主办者	cas等
学科主题	物理化学
语种	中文
ISSN号	978-7-122-00450-5
内容类型	会议论文
源URL	[http://159.226.238.44/handle/321008/112540]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
推荐引用方式 GB/T 7714	Zhao CX,Nishizawa,S.,et al. Single-nucleotide polymorphisms detection by using abasic site-containing dna duplexes[C]. 见:dalian international symposia and exhibition on chromatography. 中国. 2007-6-4.

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