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

作　　者：王学亮[1] 杨婕[2] 

机构地区：[1]菏泽学院化学与化工系,菏泽274015 [2]济宁医学院,济宁272013

出　　处：《理化检验（化学分册）》2011年第2期133-138,共6页Physical Testing and Chemical Analysis(Part B:Chemical Analysis)

基　　金：菏泽学院博士基金项目

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。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.

关 键 词：纳米金 聚硫堇 DNA 生物传感器 转基因植物 

分 类 号：O657.1[理学—分析化学]