| Budget Amount *help |
¥4,120,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥420,000)
Fiscal Year 2007: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2006: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Research Abstract |
Typing of the single nucleotide polymorphisms (SNPs) in an individual genome against sets of predetermined SNPs will be necessary for diagnosing the disease risks, predicting effects on drug treatments, and optimizing the personalized medicine. Recently, we have reported N, N' -bis(3-aminopropyl)-2, 7-diamino-1, 8-naphthyridine (DANP) is a specific stabilizer of a single cytosine and thymine bulge in duplex DNA. DANP binds to the cytosine bulge accompanied by the protonation of a nitrogen atom in the 1, 8-naphthyridine at a neutral pH. The protonated DANP (DANPH^+) bound to the cytosine emitted fluorescence at 430 nm, which was shifted by 30 nm to a longer wavelength from its unbound state. The absorption spectrum was also shifted from 360 to 400 nm upon binding to the cytosine bulge. Irradiation at 410 nm enables us selectively to excite the DANPH^+ bound to the cytosine bulge in the presence of free target DNA having C-balge. Here, we describe a new method of fluorescent SNP typing based on the C-bulge binding ligand combined with a C-balge forming DNA probes.
5' -d(ACATCCAAXCAACCAC)-3' and 17-mer C-probe 5' -(GTGGTTGYCTTGGATGT)-3', where X and Y were any nucleotide bases. When C-probe was hybridized to the target DNA, the cytosine (under lined) produced a single cytosine bulge directly neighboring to the X-Y base pair. By alternating X (A, T, C, G) in the target strand and Y (A, T, C, G, and inosine(I)) in the C-probe strand, duplexes with a single cytosine bulge flanking 20 different matched and mismatched base pairs were prepared.
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