| Project/Area Number |
14598002
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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 |
ポストゲノムのナノサイエンス
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| Research Institution | The University of Tokyo (2003)
Kyoto University (2002) |
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Principal Investigator |
OANA Hidehiro The University of Tokyo, Graduate School of Engineering, Lecturer, 大学院・工学系研究科, 講師 (20314172)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | DNA / chanize of higher order structure / phase separation in single molecule / optical tweezers / fluorescence microscopy / spermine |
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| Research Abstract |
The biochemical characteristics of lambda DNA chains in folded/unfolded states upon cleavage by the restriction enzyme ApaLI were investigated in the presence of spermine. These characteristics of DNA chains depending on their higher-order structure were studied at the single-molecule level using fluorescence microscopy. With a low concentration of spermine, lambda DNA takes a random coiled conformation and allows digestion by the enzyme, while under a high concentration of spermine, lambda DNA takes a compact folded structure and inhibits such attack. Together with comparative experiments on short oligomeric DNA, our results suggest that the transition in the higher-order structure causes on/off type switching of sensitivity to the enzyme. In addition, we proposed a noninvasive methodology for manipulating single Mb-size whole-genome DNA molecules. Cells were subjected to osmotic shock and the genorne DNA released from the burst cells was transferred to a region of higher salt concentration using optical tweezers. The trapped genome DNA exhibits a conformational transition from a condensed state to an elongated state, accompanied by the change in its environment. The applicability of optical tweezers to the on-site manipulation of giant genome DNA is suggested, i.e., lab-on-a-plate.
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