| Co-Investigator(Kenkyū-buntansha) |
NAKAO Masayuki Department of Mechanical Engineering, University of Tokyo, Professor, 工学系研究科, 教授 (90242007)
KABATA Hiroyuki Department of Mechanical Engineering, Kyoto University, Research Associatae, 工学研究科, 助手 (70293884)
KOTERA Hidetoshi Department of Mechanical Engineering, Kyoto University, Professor, 工学研究科, 教授 (20252471)
HATAMURA Yotaro Department of Mechanical Engineering, University of Tokyo, Professor, 工学系研究科, 教授 (40010863)
MATSUMOTO Kiyoshi Department of Mechanical Engineering, University of Tokyo, Professor, 情報理工学系研究科, 助教授 (10282675)
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| Research Abstract |
The purpose of this research is to develop novel methods and tools for mechanical manipulation of chromosomes, to stretch DNA fibers out of a chromosome, to analyze the structure of a chromosome, to locate a specific gene on the DNA strand, and to dissect or modify DNA (micro surgery of chromosomes). The results obtained are summarized as below.
1) A microfabricated device is developed, which enables on-demand supply of protease and its inhibitor to the object without applying excessive hydrodynamic stress. Using the device, a chromosome of higher species which appears in the division cycle of a cell is partially decomposed, and with the use of laser-manipulation methods, it has been shown that a continuous DNA fiber can be pulled out of a chromosome. It is also shown that local heating by laser is effective in partially decomposing a chromosome.
2) A method is developed to handle total chromosome of a yeast cell in a batch. The cytoskeleton of the cell is reinforced using molecular linkers to form a cage in which DNA is contained, cell membrane ruptured by adding surfactants, and FISH (Fluorescent in situ Hybridization) protocol is applied. Then the cell is placed on a solid surface, the cage structure partially destroyed, and using electroosmotic flow, DNA fiber is pulled out and stretched.
3) Electron Beam Deposition (EBD) is used to grow a tiny probe (several ten nm in diameter) on the tip of a glass needle. It is experimentally shown that aimed portion can be dissected and picked up from DNA fibers immobilized on a hydrophobic surface. The devices and methods developed in this research will be a powerful tools for bio-nanotechnology, which will bring about a new dimension of "spatial resolution" to conventional test-tube based biochemistry.
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