Inouye Masahiko, Ikeda Reona, Takase Masayoshi, Tsuri Takashi, Chiba Junya
Precursory Research for Embryonic Science and Technology, Japan Science and Technology Agency, Japan.
Proc Natl Acad Sci U S A. 2005 Aug 16;102(33):11606-10. doi: 10.1073/pnas.0502078102. Epub 2005 Aug 8.
Establishing a reliable genotyping protocol is a critical matter in post-sequence genetics. In this article, we describe a highly sensitive electrochemical detection of complementary DNAs (up to 43-mer) based on hole transport with molecular-scale, "wire-like" DNA probes. The presence of a single-base mismatch in the DNA duplexes caused a dramatic decrease in the electrochemical response. We applied this method to detect all of the possible transition and transversion SNPs and achieved "on-off"-type discrimination of fully complementary DNAs from their SNPs. Furthermore, naturally occurring polymorphisms, "hot spots" from the p53 gene, could clearly be distinguished from wild type by using our methodology.
建立可靠的基因分型方案是后测序遗传学中的关键问题。在本文中,我们描述了一种基于分子尺度的“线状”DNA探针的空穴传输对互补DNA(长达43聚体)进行高灵敏度电化学检测的方法。DNA双链体中单个碱基错配的存在会导致电化学响应急剧下降。我们应用该方法检测了所有可能的转换和颠换单核苷酸多态性(SNP),并实现了对完全互补DNA与其SNP的“开-关”型区分。此外,通过我们的方法可以清楚地将p53基因天然存在的多态性“热点”与野生型区分开来。