采用电活性标记物N-(2-乙基-二茂铁)马来酰亚胺检测与表面固定的一致性双链DNA特异性作用的p53蛋白质  

Voltammetric Detection of p53 at Electrodes Modified with Consensus Double-Stranded Oligonucleotides with N-(2-Ferrocenylethyl)maleimide

作　　者：朱旭[1] 李凯[1] 刘林[1] 王建秀[1] 刘又年[1] 

机构地区：[1]中南大学化学化工学院,长沙410083

出　　处：《化学学报》2008年第21期2379-2383,共5页Acta Chimica Sinica

基　　金：国家自然科学基金(Nos.20775093;20676153)资助项目

摘　　要：建立了电化学检测表面固定捕获的野生型p53蛋白质的方法.首先在金电极表面形成巯基化的单链DNA探针/己硫醇(HT)混合自组装膜,随后巯基化的单链DNA探针与溶液中序列匹配的靶点DNA杂交,所形成的一致性双链DNA捕获溶液中的野生型p53蛋白质.p53分子表面的半胱氨酸残基采用巯基特异性试剂N-(2-乙基-二茂铁)马来酰亚胺(Fc-Mi)进行衍生.通过检测二茂铁的电化学信号来指示p53与一致性双链DNA之间的特异性相互作用.p53蛋白质与双链DNA的键合程度取决于双链DNA的序列.该方法可检测的p53最低浓度为1.33nmol·L-1.A voltammetric method for the determination of surface-confined wild-type p53 protein has been developed. Consensus double-stranded oligonucleotide （ds-DNA） was first formed onto gold electrodes via hybridization of thiolated single-stranded DNA （ss-DNA）/hexanethiol （HT） mixed self-assembled monolayer with the complementary target DNA. The resultant consensus ds-DNA was subsequently used to capture wild-type p53 in solution. The cysteine residues on the exterior of the p53 molecules were derivatized with a thiol-specific reagent, N-（2-ferrocenylethyl）maleimide （Fc-Mi）. Well-defined voltammetric peaks that report on the interaction of p53 with the consensus ds-DNA were obtained. The level of p53 binding was found to be dependent on the sequence of the double-stranded （ds-DNA）. The present method can measure p53 concentration as low as 1.33 nmol·L^-1.

关 键 词：p53 一致性双链DNA N-(2-乙基二茂铁)马来酰亚胺 循环伏安法 

分 类 号：R73-3[医药卫生—肿瘤]