杂交及切断过程的QCM实时检测研究(英文)

电化学（中英文） ›› 2005, Vol. 11 ›› Issue (1): 32-36.

杂交及切断过程的QCM实时检测研究(英文)

朱龙章,高玉香,沈鹤柏,杨永桃,袁联群

上海师范大学生命与环境科学学院化学系,上海师范大学生命与环境科学学院化学系,上海师范大学生命与环境科学学院化学系,上海师范大学生命与环境科学学院化学系,上海师范大学生命与环境科学学院化学系上海200234 ,上海200234 ,上海200234 ,上海200234 ,上海200234

收稿日期:2005-02-28 修回日期:2005-02-28 出版日期:2005-02-28 发布日期:2005-02-28

In-situ Measurements of Single-stranded DNA Hybridization and Cleavage by Quartz Crystal Microbalance

ZHU Long-zhang, GAO Yu-xiang, SHEN He-bai~*, YANG Yong-tao, YUAN Lian-qun

(Dept. of Chem., Shanghai Normal University, Shanghai 200234, China

Received:2005-02-28 Revised:2005-02-28 Published:2005-02-28 Online:2005-02-28

摘要/Abstract

摘要： 采用自组装技术,将 5′端标记有巯基的 20-merODN(oligo 1)以金硫键形式牢固结合在 7. 995MHz的AT-切石英晶体的镀金表面,然后由石英晶体微天平实时检测了与碱基序列互补的 10 merODN (oligo 2)和 8 merODN(oligo 3)的杂交,同时还研究了稀土金属铈离子在温和条件下对DNA的水解切断作用.结果表明:应用QCM方法可能实时检测DNA的固定和杂交,Ce(IV)能随机切断单链DNA;但不能切断杂交形成的双链DNA,因此可利用杂交保护的方法对单链DNA实行定位切断.

关键词: 石英晶体微天平, 单链脱氧核糖核酸

Abstract: The use of quartz crystal microbalance (QCM) for in-situ monitoring the immobilization of single-stranded DNA(oligo-1), marked with mercaptol group at 5′ end, on the surface of gold-filled 7.995 MHz AT-cut quartz crystal by Au-S bond with self-assemble technique is reported. And then the hybridization of ssDNA with the complementary 10-mer ODN(oligo-2) and 8-mer ODN(oligo-3) is described. Moreover, the QCM has been employed to analyze the DNA cleavage by cerium(IV) ions under moderate conditions. The results showed that the QCM, which is capable of sensitive measurement, was able to in-situ investigate immobilization, hybridization and cleavage of ssDNA. And the cerium(IV) ions had no cleavage for the double-stranded DNA, but it could hydrolyze single-stranded DNA. So the hydrolytic cleavage of ssDNA at the specific site could be realized by the protection of the hybridization.

Key words: Quartz crystal microbalance, Qligodeoxynucleotide, Hybridization, Hydrolytic cleavage

中图分类号:

Q503

朱龙章,高玉香,沈鹤柏,杨永桃,袁联群. 杂交及切断过程的QCM实时检测研究(英文)[J]. 电化学（中英文）, 2005, 11(1): 32-36.

ZHU Long-zhang, GAO Yu-xiang, SHEN He-bai~*, YANG Yong-tao, YUAN Lian-qun. In-situ Measurements of Single-stranded DNA Hybridization and Cleavage by Quartz Crystal Microbalance[J]. Journal of Electrochemistry, 2005, 11(1): 32-36.

导出引用管理器 EndNote|Ris|BibTeX

链接本文: https://electrochem.xmu.edu.cn/CN/

https://electrochem.xmu.edu.cn/CN/Y2005/V11/I1/32

参考文献

[1]　PatolskyF,LichtensteinA,WillnerL, etal.Amodi fiedmicrogravimetricquartz crystalmicrobalanceassayofDNAusingoligonucleotidefunctionalizedliposomesorbiotinylateliposomes[J].J.Am.Chem.Soc.,2002,122:418~419. [2]　AberlF,WolfH.HIVserologyusingpiezoelectricim munosensors[J].SensorsandActuatorsB,1994,18~19:271~275. [3]　ZhouXichun,HuangLiqun,SamfongYaoyi.Micro gravimetricDNAsensorbasedonquartzcrystalmi crobalance:comparisonofoligonucleotideimmobilizationmethodsandtheapplicationingeneticdiagnosis[J].Biosensor&Bioelectronics,2001,16:85~91. [4]　MoXiangTao,ZhouYiPing,LeiHan, etal.Mi crobalanceDNAprobemethodforthedetectionofspecificbacteriainwater[J].EnzymeandMicrobialTech nology,2002,30:583~589. [5]　YoshioOkahata,KenichiNiikura,YukioSugiura,etal.Kineticstudyofsequence specificbindingofGCN4 bZ IPpeptidestoDNAstrandsimmobilizedona27MHzquartzcrystalmicrobalance[J].Biochemistr.,1998,37:5666~5672. [6]　ShenHebai,Xiajingfen,YangHaifeng, etal.Hydrol ysisofoligodeoxynucleotidephosphodiesterlinkages[J].ScienceinChina(seriesB),2001,44,(2):169~174. [7]　FrankCaruso,ElkeRodda,NeilFurlongD,etal.QuartzcrystalmicrobalancestudyofDNAimmobilizationandhybridizationfornucleicacidsensordevelopment[J].Anal. chem.,1997,69:2043~2049. [8]　KeikoIto,KojiHashimoto,YoshioIshimori.Quantita tiveanalysisforsolid phasehybridizationreactionandbindingreactionofDNAbindertohybridsusingaquartzcrystalmicrobalance[J].AnalyticaChimicaActa,1996,327:29~35. [9]　YoshioOkahata,MasanoriKawase,KenichiNikura, etal.KineticmeasurementsofDNAhybridizationonanoligonucleotideimmobilized27MHzquartzcrystalmi crobalance[J].Anal.Chem.,1998,70:1288~1296. [10]　BrahamUlman,JungFKang,YitzhakShnidman, etal.Self assembledmonolayersofrigidthiols[J].Re viewsinMolecularBiotechnology,2000,74:175~188. [11]　SunXingyan,HePinggang,LiuShenghui, etal.Im mobilizationofsingle strandeddeoxyribonucleicacidongoldelectrodewithself assembledaminoethanethiolmonolayerforDNAelectrochemicalsensorapplications[J].Talanta,1998,47:487~495. [12]　KomiyamaM,TakedaN,ShigekawaH.HydrolysisofDNAandRNAbylanthanideionsMechanisticstudiesleadingtonewapplications[J].ChemicalCommunica tions,1999,16:1443~1551. [13]　IgawaT,SumaokaJ,KomiyamaM.Hydrolysisofoli gonucleotidesbyhomogeneousCe(IV)/EDTAcomplex[J].Chemistryletters,2000:356~357. [14]　ShenHebai,ZhouWenjun,YangYongtao, etal.Hy drolyticscissionofDNAbyceriumoligoDNAhybridwithsequence specificity[J].ScienceinChina(seriesB),.2004,(1):75.