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以富勒烯C60,L-苏氨酸及对苯二甲醛为原料,在氮气保护下反应得到含醛基官能团的2-(4-醛基苯基)-5-(1-羟乙基)富勒烯吡咯烷衍生物(C60-CHO).将该材料修饰于玻碳电极表面,并利用醛基与氨基之间温和、高效的缩合反应,将5’-氨基修饰的寡聚核苷酸共价固定到了C60-CHO修饰的玻碳电极表面,构建了一种新型的电化学DNA传感器.以[Fe(CN)6]3?/4?为电活性探针,采用电化学阻抗法对转基因植物CaMV35S启动子基因特征片段进行检测.实验结果表明,杂交前后的电子传递电阻差值(ΔRet)与目标序列浓度对数(lg CS2)在1.0×10-13～1.0×10-9 mol/L浓度范围内呈良好的线性关系,线性回归方程为ΔRet/(103?)=3.471 lg(CS2/mol/L)+50.425(r=0.9977),检测限为1.5×10?14 mol/L.杂交特异性实验进一步表明该传感器对完全互补、碱基错配和非互补序列具有良好的识别能力.
The reaction of fullerene C60, L-threonine and terephthalaldehyde under the protection of nitrogen afforded 2- (4-allylphenyl) -5- (1-hydroxyethyl) Fullerene pyrrolidine derivative (C60-CHO). This material was modified on the surface of a glassy carbon electrode and the 5 ’-amino-modified oligonucleotide was modified by a mild and efficient condensation reaction between the aldehyde group and the amino group A new type of electrochemical DNA sensor was constructed by covalently immobilizing C60-CHO on the surface of glassy carbon electrode. Electrochemical impedance was used to detect the electrochemical reaction of [Fe (CN) 6] 3? / 4? The results showed that the difference between the electron transfer resistance (ΔRet) and the target sequence concentration (lg CS2) before and after hybridization was 1.0 × 10-13 ~ 1.0 × 10-9 mol / The linear regression equation was ΔRet / (103?) = 3.471 lg (CS2 / mol / L) +50.425 (r = 0.9977) with a detection limit of 1.5 × 10 ~ 14 mol / L. Hybridization specificity experiments further demonstrate that the sensor has good recognition of perfectly complementary, base mismatched and non-complementary sequences.