| Project/Area Number |
23510139
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Nanomaterials/Nanobioscience
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| Research Institution | The Institute of Physical and Chemical Research |
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Principal Investigator |
AOKI Hiroyoshi 独立行政法人理化学研究所, 光量子工学研究領域, 研究員 (50518636)
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| Co-Investigator(Kenkyū-buntansha) |
IKE Fumio 独立行政法人理化学研究所, バイオリソースセンター, 専任研究員 (40183157)
SHINTANI Masaki 静岡大学, 工学(系)研究科(研究院), 准教授 (20572647)
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| Project Period (FY) |
2011-04-28 – 2015-03-31
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| Project Status |
Completed (Fiscal Year 2014)
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| Budget Amount *help |
¥5,720,000 (Direct Cost: ¥4,400,000、Indirect Cost: ¥1,320,000)
Fiscal Year 2013: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000)
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| Keywords | 難培養微生物 / アガロースフィルム / 一菌体分離 / 全ゲノム増幅 / レーザーマイクロダイセクション / マイクロ流路 / ナノファイバー / 一菌体ゲノム増幅 / マイクロ流体チップ |
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| Outline of Final Research Achievements |
To analyze yet-cultured microbes (YCM) in the environment, device and technique were studied to manually isolate single cell.
(1) For manually isolation, a slow-flow focusing microchannel was developed using electrophoresis and hydrostatic pressure pumps. The pumps reduced the flow rates at less than 1/10 of conventional pump, enabling operator to recognize flowing cells.
(2) To prevent contamination, single microbe was isolated by embedding in agarose. An agarose film was prepared from a gel containing E. coli, laser absorbent, and hydrophilic polymer (PVA) by drying. From the agarose film, a small piece (approx. 100 x 100 x 20 um) containing single cell was dissected by laser microdissection. After the piece was swelled in buffer by dissolving PVA, the E. coli genomic DNA was specifically amplified in the swelled agarose by Phi 29 DNA polymerase. Thus, the developed device and technology will provide unique tools for symbiotic genomics of specific environment-colonized YUM.
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