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学科主题: 药学
题名:
Gold nanoparticle enhanced fluorescence anisotropy for the assay of single nucleotide polymorphisms (SNPs) based on toehold-mediated strand-displacement reaction
作者: Wang, Xinyi1,2,3; Zou, Mingjian2; Huang, Hongduan2; Ren, Yuqian2; Li, Limei1; Yang, Xiaoda3; Li, Na2
关键词: Fluorescence anisotropy ; Gold nanoparticles ; Single nucleotide polymorphism ; Strand-displacement
刊名: BIOSENSORS & BIOELECTRONICS
发表日期: 2013-03-15
DOI: 10.1016/j.bios.2012.09.023
卷: 41, 页:569-575
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Biophysics ; Biotechnology & Applied Microbiology ; Chemistry, Analytical ; Electrochemistry ; Nanoscience & Nanotechnology
研究领域[WOS]: Biophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics
关键词[WOS]: NEAREST-NEIGHBOR THERMODYNAMICS ; DOT-T MISMATCHES ; DNA HYBRIDIZATION ; INVASIVE CLEAVAGE ; MASS-SPECTROMETRY ; PROBES ; POLARIZATION ; DEPENDENCE ; MUTATIONS ; PLATFORM
英文摘要:

We developed a highly differentiating, homogeneous gold nanoparticle (AuNP) enhanced fluorescence anisotropic method for single nucleotide polymorphism (SNP) detection at nanomolar level using toehold-mediated strand-displacement reaction. The template strand, containing a toehold domain with an allele-specific site, was immobilized on the surface of AuNPs, and the solution fluorescence anisotropy was markedly enhanced when the fluorescein-labeled blocking DNA was attached to the AuNP via hybridization. Strand-displacement by the target ssDNA strand resulted in detachment of fluorescein-labeled DNA from AuNPs, and thus decreased fluorescence anisotropy. The drastic kinetic difference in strand-displacement from toehold design was used to distinguish between the perfectly matched and the single-base mismatched strands. Free energy changes were calculated to elucidate the dependence of the differentiation ability on the mutation site in the toehold region. A solid negative signal change can be obtained for single-base mismatched strand in the dynamic range of the calibration curve, and a more than 10-fold signal difference can still be observed in a mixed solution containing 100 times the single-base mismatched strand, indicating the good specificity of the method. This proposed method can be performed with a standard spectrofluorimeter in a homogeneous and cost-effective manner, and has the potential to be extended to the application of fluorescence anisotropy method of SNP detection. (C) 2012 Elsevier B.V. All rights reserved.

语种: 英语
所属项目编号: 20975004 ; 21035005 ; L2011108
项目资助者: National Natural Science Foundation of Chin ; Instrumental Analysis Fund of Peking University ; Education Department of Liaoning Province
WOS记录号: WOS:000314191300085
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/63331
Appears in Collections:北京大学药学院_化学生物学系_期刊论文

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作者单位: 1.Shenyang Agr Univ, Coll Sci, Shenyang 110161, Peoples R China
2.Peking Univ, Key Lab Bioorgan Chem & Mol Engn, Coll Chem & Mol Engn, BNLMS,Minist Educ, Beijing 100871, Peoples R China
3.Peking Univ, Dept Biol Chem, Sch Pharmaceut Sci, Beijing 100083, Peoples R China

Recommended Citation:
Wang, Xinyi,Zou, Mingjian,Huang, Hongduan,et al. Gold nanoparticle enhanced fluorescence anisotropy for the assay of single nucleotide polymorphisms (SNPs) based on toehold-mediated strand-displacement reaction[J]. BIOSENSORS & BIOELECTRONICS,2013,41:569-575.
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