用荧光探针签别CYP2C9*3基因

用一种基激复合物荧光探针系统来签别碱基错配的CYP2C9*3基因，该系统选择两个分开的与靶点碱基相对应的12碱基荧光标记的寡核苷酸作为探针，分别对24碱基、47碱基、质粒(3165bp)的靶点CYP2C9基因和CYP2C9*3基因进行杂交配对，结果该基激复合物荧光探针系统能有效签别各种长度的CYP2C9基因和CYP2C9*3基因，背景干扰很低，灵敏度高，可尝试用于其他基因型的遗传多态性的签别。

A Fluorescent-Probe Detecting CYP2C9*3 DNA

The mismatched CYP2C9*3 DNA was detected by a exciplex fluorescent-probe system. The exciplex fluorescent-probe model system comprises of two 12-mer oligonucleotides fluorescent-probes, complementary to neighbouring sites of a 24(47) -mer DNA target and plasmid target, each equipped with moieties able to form an exciplex on correct, contiguous hybridization. Very similar results were obtained between the 24-mer target and the 47-mer target when WT and MT systems were detected by the exciplex fluorescent-probe. Exciplex bands at 505 nm were found for both 24(47)-mer WT and 24(47)-mer MT-targets at 5 ℃ , but were not distinctive enough to distinguish 24(47)-mer WT-target and 24(47)-mer MT-target. However the experiments were carried out at Tm, the exciplex band disappeared almost completely for 24(47)-mer MT-target system like control system, and there was still a strong exciplex band for the 24(47)-mer WT target system. Exciplex peaks at 505 nm were seen for the WT circular plasmids system, but not for MT circular plasmids. Therefore, mismatches of CYP2C9*3 DNA can be effectively detected by this exciplex construct, giving potential for single nucleotide polymorphism detection.