2007 Fiscal Year Final Research Report Summary
Fluorescent Determination of SNPs by C-Bulge binding ligand
| Project/Area Number |
18550071
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Analytical chemistry
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| Research Institution | Osaka University |
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Principal Investigator |
TAKEI Fumie Osaka University, The institute of Scientific and Industrial Research, Research Associates (30252711)
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| Co-Investigator(Kenkyū-buntansha) |
NAKATANI Kazuhiko Osaka University, The institute of Scientific and Industrial Research, Professor (70237303)
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| Project Period (FY) |
2006 – 2007
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| Keywords | SNP / Fluorescent molecule / DNA / Cvtosine bulge |
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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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[Journal Article] Allele Specific C-bulge Probes with One Unique Fluorescent Molecule Discriminate the Single Nucleotide Difference in DNA2007
Author(s)
F., Takei, H., Suda, M., Hagihara, J., Zhang, A., Kobori, K., Nakatani
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Journal Title
Chem. -Eur. J 13
Pages: 4452-4457
Description
「研究成果報告書概要(欧文)」より
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[Presentation] Fluorescent Determination of SNPs using N, N'-bis(3-aminopropyl)-2,7-diamino-1,8-naphthyridine2007
Author(s)
F., Takei, M., Hagihara, J., Zhang, K., Nakatani
Organizer
17th International Roundtable on Nucleosides, Nucleotides and Nucleic Acids(IRTXVII), 3rd Spring
Place of Presentation
Switzerland
Year and Date
20070000
Description
「研究成果報告書概要(欧文)」より
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