Mukumoto Kosuke, Watanabe Sadayoshi, Nojima Takahiko, Waki Michinori, Takenaka Shigeori
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, Japan.
Anal Sci. 2007 Jun;23(6):645-9. doi: 10.2116/analsci.23.645.
Ferrocenylcarbodiimide (1), which is known to react with a guanine (G) or thymine (T) base of single stranded DNA, was allowed to react with DNA duplex having a single mismatched base pair of G-T, T-T, or T-cytosine (C). Electrophoreograms of the reaction mixture showed that 1 could react with G or T base of the mismatched sites on the DNA duplex. However, 1 also reacted with the G base of the terminal site on the DNA duplex. This showed that 1 can react with an unpaired base or unstable base pair such as a terminal or mismatched base on the DNA duplex. Electrochemical mismatch detection could be achieved after hybridization of the ferrocenylated mismatched DNA duplex with a selected DNA probe-immobilized electrode. These results revealed that 1 has a potentiality of serving as a labeling reagent of mismatched bases on the DNA duplex, which is important in the search for heterozygous single nucleotide polymorphisms (SNPs).
已知二茂铁基碳二亚胺（1）能与单链DNA的鸟嘌呤（G）或胸腺嘧啶（T）碱基发生反应，现将其与具有G-T、T-T或T-胞嘧啶（C）单碱基错配的DNA双链体进行反应。反应混合物的电泳图谱表明，1能与DNA双链体错配位点上的G或T碱基发生反应。然而，1也会与DNA双链体末端位点的G碱基发生反应。这表明1能与未配对碱基或不稳定碱基对发生反应，如DNA双链体上的末端或错配碱基。在将二茂铁基化的错配DNA双链体与选定的固定有DNA探针的电极杂交后，可实现电化学错配检测。这些结果表明，1有潜力作为DNA双链体上错配碱基的标记试剂，这在寻找杂合单核苷酸多态性（SNP）方面具有重要意义。
