Cho Minseon, Lee Sohyun, Han Se-Young, Park Jin-Young, Rahman Md Aminur, Shim Yoon-Bo, Ban Changill
Department of Chemistry, Pohang University of Science and Technology, Pohang, Gyungbuk, 790-784, South Korea.
Nucleic Acids Res. 2006 Jun 13;34(10):e75. doi: 10.1093/nar/gkl364. Print 2006.
A direct and label-free electrochemical biosensor for the detection of the protein-mismatched DNA interaction was designed using immobilized N-terminal histidine tagged Escherichia coli. MutS on a Ni-NTA coated Au electrode. General electrochemical methods, cyclic voltammetry (CV), electrochemical quartz crystal microbalance (EQCM) and impedance spectroscopy, were used to ascertain the binding affinity of mismatched DNAs to the MutS probe. The direct results of CV and impedance clearly reveal that the interaction of MutS with the CC heteroduplex was much stronger than that with AT homoduplex, which was not differentiated in previous results (GT > CT > CC approximately AT) of a gel mobility shift assay. The EQCM technique was also able to quantitatively analyze MutS affinity to heteroduplexes.
利用固定在镍-氮三乙酸(Ni-NTA)修饰金电极上的N端组氨酸标记的大肠杆菌MutS,设计了一种用于检测蛋白质-错配DNA相互作用的直接且无标记的电化学生物传感器。采用常规电化学方法,即循环伏安法(CV)、电化学石英晶体微天平(EQCM)和阻抗谱,来确定错配DNA与MutS探针的结合亲和力。CV和阻抗的直接结果清楚地表明,MutS与CC异源双链体的相互作用比与AT同源双链体的相互作用强得多,而在先前凝胶迁移率变动分析的结果(GT > CT > CC约等于AT)中并未区分这种差异。EQCM技术也能够定量分析MutS对异源双链体的亲和力。