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DNA电化学生物传感器在转基因植物特定序列基因检测中的应用
          
Application of DNA Electrochemical Biosensors to Detection of Specific Genetic Squences in Transgenetic Plants

摘    要
在玻碳电极(GCE)上采用循环伏安法电聚合硫堇(PTh)得到PTh/GCE修饰电极,并利用聚硫堇层共价结合和静电作用吸附金纳米粒子(AuNP′s)制得AuNP′s/PTh/GCE修饰电极。然后通过将ss-DNA/AuNP′s/PTh修饰电极置于cDNA杂交液中,于42 ℃杂交制得ds-DNA/AuNP′s/PTh修饰玻碳电极,实现了脱氧核糖核酸(DNA)探针在AuNP′s/PTh修饰的玻碳电极上的固定,制得DNA电化学生物传感器。在[Fe(CN)6]3-/4-溶液中采用微分脉冲伏安法(DPV)及交流阻抗谱技术(EIS)对DNA的固定和杂交进行了表征。试验结果表明:在1.0×10-10~1.0×10-6mol·L-1的浓度范围内,该传感器可对转基因植物外源基因草丁膦乙酰转移酶基因(PAT基因)片段进行检测,检出限(3s)为3.2×10-11mol·L-1。
标    签 纳米金   聚硫堇   DNA   生物传感器   转基因植物   Nanogold   Polythionine   DNA   Biosensor   Transgenetic plants  
 
Abstract
The glassy carbon electrode (GCE) was modified with polythionine (PTh) to form PTh/GCE by electropolymerization of thionine with cyclic voltammetry, and nanoparticles of gold were adhered to the PTh/GCE through coordination and electrostatic action of the PTh layer to form the modified electrode of AuNP′s/PTh/GCE. Finally, DNA probe was immobilized onto the AuNP′s/PTh/GCE through the interaction between ss-DNA and nano-gold to form ss-DNA/AuNP′s/PTh/GCE, which was further hydridized in cDNA solution at 42 ℃ to form ds-DNA/AuNP′s/PTh/GCE. The immobilization and hydridization of DNA were characterized by differential pulse voltammetry and AC impedance spectroscopy in a solution of [Fe(CN)6]3-/4-. It was found that linear relationship between values of detection signals (ΔRet) and concentration of the 20-base PAT genetic sequence was obtained in the range of 1.0×10-10-1.0×10-6mol·L-1. Detection limit (3s) of this biosensor was found to be 3.2×10-11mol·L-1.

中图分类号 O657.1

 
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基金项目 菏泽学院博士基金项目

收稿日期 2009/11/13

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备注王学亮(1977-),男,山东东明人,博士,主要从事生命电分析化学研究。

引用该论文: WANG Xue-liang,YANG Jie. Application of DNA Electrochemical Biosensors to Detection of Specific Genetic Squences in Transgenetic Plants[J]. Physical Testing and Chemical Analysis part B:Chemical Analysis, 2011, 47(2): 133~138
王学亮,杨婕. DNA电化学生物传感器在转基因植物特定序列基因检测中的应用[J]. 理化检验-化学分册, 2011, 47(2): 133~138


被引情况:


【1】汪涛,王朝霞,王学亮,郁章玉, "基于血红蛋白/金纳米粒子/聚二烯丙基二甲基氯化铵-多壁碳纳米管生物传感器测定过氧化氢",理化检验-化学分册 49, 577-579(2013)

【2】焦翠玲,王学亮,同元辉,郁章玉, "壳聚糖/DNA/血红蛋白/多壁碳纳米管复合膜修饰玻碳电极的制备及其电化学行为的研究",理化检验-化学分册 49, 15-18(2013)



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