Efficient polymerase chain reaction from single DNA molecules
| Project/Area Number |
18K14260
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| Research Category |
Grant-in-Aid for Early-Career Scientists
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| Allocation Type | Multi-year Fund |
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| Review Section |
Basic Section 34020:Analytical chemistry-related
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| Research Institution | Japan Agency for Marine-Earth Science and Technology |
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Principal Investigator |
Zhang Yi 国立研究開発法人海洋研究開発機構, 超先鋭研究開発部門(超先鋭研究プログラム), 研究員 (40795358)
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| Project Period (FY) |
2018-04-01 – 2021-03-31
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| Project Status |
Completed (Fiscal Year 2020)
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| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2020: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2018: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
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| Keywords | High throughput / Microfabrication / Single molecule / Enzyme / Sensitivity / Poisson statistics / Image processing / Screening / Restriction enzyme / Electrophoresis / Digital counting / Droplet / Biosensor / Cell-free / Directed evolution / Synthetic biology / PCR / Protein synthesis / Micromanipulation / DNA / Bioreactor / Microdevice |
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| Outline of Final Research Achievements |
We cannot study what we cannot measure. This simple philosophy is the basic driving force for analytical chemistry to move forward. This Early-Career Scientists study was also planned and implemented based on this general interest. My research platform features high throughput and deals with a massive number of single DNA molecules, which enabled: (1) cell-free protein synthesis from every single DNA molecule; (2) genotype-phenotype linkage at the single DNA molecule; (3) sorting and amplification for single DNA molecules; and (4) amplification-free single DNA molecule detection. During the period of this research, I succeeded in obtaining the world's most active enzyme capable of hydrolyzing phosphate esters, establishing the novel digital high-throughput screening theory, and developing the world's most sensitive method capable of evaluating the performance of any DNA-cleaving enzymes.
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| Academic Significance and Societal Importance of the Research Achievements |
The massively parallel, highly uniform, and individually addressable femtoliter droplet array technology offered a high-end standard and new possibilities for future diverse bioanalytical applications. The integrated system also stressed the significance of the multidisciplinary approach.
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Report
(4 results)
Research Products
(17 results)
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[Journal Article] Multiplexed single-molecule enzyme activity analysis for counting disease-related proteins in biological samples2020
Author(s)
Shingo Sakamoto, Toru Komatsu, Rikiya Watanabe, Yi Zhang, Taiki Inoue, Mitsuyasu Kawaguchi, Hidehiko Nakagawa, Takaaki Ueno, Takuji Okusaka, Kazufumi Honda, Hiroyuki Noji, Yasuteru Urano
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Journal Title
Science Advances
Volume: 6
Issue: 11
DOI
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Peer Reviewed / Open Access
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[Presentation] １分子酵素活性プロファイリングによる疾患関連酵素の超高感度検出2019
Author(s)
坂本眞伍, 小松徹, 渡邉力也, 張翼, 井上大輝, 川口充康, 中川秀彦, 植野高章, 奥坂拓志, 本田一文, 野地博行, 浦野泰照
Organizer
第92回日本生化学会大会
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