[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 degrees C, 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.
Li
Qing-Yong
QY
Key Laboratory of Forest Plant Ecology (Northeast Forestry University), Ministry of Education, Harbin 150040, China. liqingyong2004@yahoo.com.cn
Qu
Zhen-Huan
ZH
Zu
Yuan-Gang
YG
Fu
Yu-Jie
YJ
D065729:Cytochrome P-450 CYP2C9 / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D009693:Nucleic Acid Hybridization; D010957:Plasmids
D015342:DNA Probes; D005456:Fluorescent Dyes; D004247:DNA; D065729:Cytochrome P-450 CYP2C9