| Project/Area Number |
16350090
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Chemistry related to living body
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| Research Institution | Kanagawa University (2005-2007)
Tokyo Metropolitan University (2004) |
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Principal Investigator |
ONO Akira Kanagawa University, Faculty of Engineering, Professor (10183253)
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| Co-Investigator(Kenkyū-buntansha) |
OKAMOTO Itaru Kanagawa University, Faculty of Engineering, assistant Professor (40460133)
TASHIRO Mitsuru Meisei University, School of Science and Engineering, Professor (40315750)
三宅 洋子 首都大学東京, 都市教養学部, 助手 (40244412)
市村 徹 首都大学東京, 都市教養学部, 准教授 (50213012)
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| Project Period (FY) |
2004 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥14,010,000 (Direct Cost: ¥13,200,000、Indirect Cost: ¥810,000)
Fiscal Year 2007: ¥3,510,000 (Direct Cost: ¥2,700,000、Indirect Cost: ¥810,000)
Fiscal Year 2006: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2005: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 2004: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | SNP / single nucleotide polymorphism / DNA / metal ions / DNA duplex / thermal stability / mercury / silver / 水銀イオン / 銀イオン / Hg(II) / Ag(I) / 熱量測定 |
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| Research Abstract |
The goal of the research is developing a new efficient methods for detecting single-nucleotide-polymorphism (SNP) using a phenomenon, certain metal ions highly selectively bind to miss base pairs in DNA duplexes, recently found in our laboratory. Mercury(II) and silver(I) ions highly selectively bind to thymine-thymine (T-T) pair and cytosine-cytosine (C-C) pair in DNA duplexes, respectively, and consequently the duplexes containing the miss base pairs are stabilized in the presence of the metal ions. The phenomenon is totally different form the existed methods in which metal ions are captured by artificial base pairs in DNA duplexes. This finding that the natural bases selectively bind to the certain metal ions is scientifically interesting, as well as this can be used for developing new SNP typing methods.
In this study, first of all, we examined a capability that a SNP typing method can be developed by detecting the increased thermal stability of a duplex in the presence of the metal ions. We used the isothermal titration calorimetric experiments, and some results were published. Also, we tried to detect SNP from the fluorescence emission intensity differences in the absence and in the presence of the metal ions.
Beside the application studies, we thermodynamically studied binding processes of the metal ions and the base pairs. We tried to clarify the structure of the metal ion binding sites, and NMR method was efficient for Hg(II)-DNA and Ag(I)-DNA complexes.
We have searched new metal ion-base pair combinations in which metal ions selectively and tightly bind to base pairs and DNA duplexes are stabilized in the presence of metal ions. However, such metal ion-base pair combinations were not found. Also, we searched new metal mediated base pairs in which the natural bases and artificial bases are tightly connected by metal ions, and the study has been in progress.
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