金电极自组装基因分子膜的制备及电化学检测  

Preparation and electrochemical determination of gold electrode modified with thiolated capture probe through self-assembling

作　　者：黎媛萍[1,2,3] 曾光明[1,3] 汤琳[1,3] 陈耀宁[1,3] 章毅[1,3] 刘灿[1,3] 李贞[1,3] 庞娅[1,3] 

机构地区：[1]湖南大学环境科学与工程学院,长沙410082 [2]湖南城市学院化学与环境工程学院,益阳413000 [3]湖南大学环境生物与控制教育部重点实验室,长沙410082

出　　处：《中国有色金属学报》2013年第5期1345-1350,共6页The Chinese Journal of Nonferrous Metals

基　　金：教育部长江学者和创新团队发展计划资助项目(IRT0719);教育部新世纪优秀人才支持计划资助项目(NCET-11-0129);国家自然科学基金资助项目(50608029;50808072;50978088);湖南大学中央高校基本科研业务费专项资金资助项目;湖南省优秀博士学位论文科研项目

摘　　要：采用分子自组装技术制备巯基修饰的基因分子膜修饰金电极,用于竞争式杂交检测黄孢原毛平革菌木素过氧化物酶编码基因。通过差分脉冲伏安法、循环伏安法、交流阻抗法和电流—时间曲线法优化自组装时间和信号探针的最佳响应浓度,研究目标基因的线性检测范围和再生性能。结果表明:修饰金电极最优自组装时间为15 h,信号探针的最佳响应浓度为0.51×10-6 mol/L,目标基因的线性检测范围为7.51×10-12-1.05×10-9 mol/L,检测下限为7.51×10-13 mol/L。该修饰电极具有良好的再生性能。A gold electrode with thiolated capture probe through self-assembling based on the competitive hybridization for detection of target sequence of lignin peroxidase （lip） gene of Phanerochaete chrysosporium was developed. Following hybridizations with competitively hybridized with the target nucleic acid and biotinylated response probe, streptavidin-horseradish peroxidase （HRP） conjugate was applied to the electrode. The electrochemical behavior was analyzed by cyclic voltammetry, electrochemical impedance spectroscopy, current--time curve and differential pulse voltammetry. The results show that the best time for self-assembling is 15 h, and the optimum concentration of response probe is 0.51×10^-6 mol/L. A good linear correlation between the current and the concentration of the target nucleic acid concentration is found in the concentration range from 7.51×10^-12 mol/L to 1.05×10^-9 mol/L. The detection limit is 7.51×10^-13 mol/L. The modified electrode exhibits high sensitivity, precision, stability and reproducibility.

关 键 词：修饰金电极 自组装单分子膜 基因 竞争杂交 电化学检测 

分 类 号：O657.1[理学—分析化学]