基于DNA聚合酶I的新型电化学传感器及其胰腺癌K-ras基因点突变检测

doi:10.13208/j.electrochem.140725

电化学（中英文） ›› 2015, Vol. 21 ›› Issue (1): 72-77. doi: 10.13208/j.electrochem.140725

基于DNA聚合酶I的新型电化学传感器及其胰腺癌K-ras基因点突变检测

林丽清，赵成飞，蒋周倩，翁少煌，谢晓兰，林新华^*

福建医科大学药学院，福建福州 350004

收稿日期:2014-07-25 修回日期:2014-09-26 出版日期:2015-02-28 发布日期:2014-10-08
通讯作者: 林新华 E-mail:xinhua63@163.com
基金资助:
国家863计划（No. 2012AA022604）、国家自然科学基金项目（No. 21405016，No. 21275028）资助

Novel DNA Electrochemical Biosensor for K-Ras Point Mutation Detection Based on DNA Polymerase I

LIN Li-qing, ZHAO Cheng-fei, JIANG Zhou-qian, WENG Shao-huang, XIE Xiao-lan, Lin Xin-hua^*

Faculty of Pharmacy, Fujian Medical University, Fuzhou 350004, China

Received:2014-07-25 Revised:2014-09-26 Published:2015-02-28 Online:2014-10-08
Contact: Lin Xin-hua E-mail:xinhua63@163.com

摘要/Abstract

摘要： 本文构建DNA聚合酶I的新型DNA电化学传感器，将捕获探针通过Au—S键固定于Au基底表面，与互补靶序列杂交至点突变前一个碱基，通过DNA聚合酶Ⅰ将dUTP-biotin连接在目标DNA的检测位点，再与avidin-HRP反应，而后测定在TMB溶液中的电化学特性. 结果表明，DNA电化学传感电极的检测电流值与K-ras突变型基因浓度（1.0×10^-15 ~ 1.0×10^-10 mol·L^-1）对数呈良好的线性关系，且灵敏度高，特异性较佳.

关键词: DNA聚合酶I, K-ras基因点突变, DNA电化学传感器, 电流时间曲线

Abstract: In this article, we developed a novel DNA electrochemical sensor for obtaining a current-time curve based on DNA polymerase I. The capture probes were fixed on the Au electrode by Au interacting with -SH, and then hybridized with the K-ras mutant target DNA as far as one base before the mutation site. After hybridizing the K-ras mutant or wild target DNA, the resulting assembly reacted with the dUTP-biotin in the presence of DNA polymerase I. Then avidin-HRP combined the dUTP-biotin on the electrode. The proposed electrode was applied for detection in TMB solution by electrochemical technology. The experimental results showed that the detection current of the proposed DNA electrochemical sensor is proportional to the logarithmic K-ras mutant DNA concentration (1.0×10^-15 to 1.0×10^-10 mol·L^-1). It had high sensitivity and good selectivity.

Key words: DNA polymerase I, K-ras point mutant gene, DNA electrochemical sensor, current-time curve

中图分类号:

O646

林丽清，赵成飞，蒋周倩，翁少煌，谢晓兰，林新华*. 基于DNA聚合酶I的新型电化学传感器及其胰腺癌K-ras基因点突变检测[J]. 电化学（中英文）, 2015, 21(1): 72-77.

LIN Li-qing, ZHAO Cheng-fei, JIANG Zhou-qian, WENG Shao-huang, XIE Xiao-lan, Lin Xin-hua*. Novel DNA Electrochemical Biosensor for K-Ras Point Mutation Detection Based on DNA Polymerase I[J]. Journal of Electrochemistry, 2015, 21(1): 72-77.

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基于DNA聚合酶I的新型电化学传感器及其胰腺癌K-ras基因点突变检测

Novel DNA Electrochemical Biosensor for K-Ras Point Mutation Detection Based on DNA Polymerase I

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