2019 Fiscal Year Research-status Report
Efficient polymerase chain reaction from single DNA molecules
| Project/Area Number |
18K14260
|
|---|
| Research Institution | Japan Agency for Marine-Earth Science and Technology |
|---|
Principal Investigator |
張 翼 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)
|
|---|
| Project Period (FY) |
2018-04-01 – 2021-03-31
|
| Keywords | Cell-free / Screening / Directed evolution / Synthetic biology / Microfabrication |
|---|
| Outline of Annual Research Achievements |
I optimized the single-molecule DNA recovery platform and applied this platform for an accurate high-throughput enzyme screening independent of cell culture (Yi Zhang et al., Science Advances, 2019). In this work, I proposed a novel strategy based on Poisson statistics to enable rapid and efficient acquisition of enzyme mutants from just a few clones. This is a new digitalized biotechnology in parallel with digital polymerase chain reaction (digital PCR) and digital enzyme-linked immunosorbent assay (digital ELISA). To better spread this new research tool, I accepted an invitation from a video journal and offered a detailed protocol regarding my original technology (Yi Zhang et al., JoVE, 2020). The optimized protocol makes the novel research platform accessible to everyone who has standard lithographic facilities and a conventional fluorescence microscope in their own place. Besides, the extension of my technology toward the performance assessment of DNA-related enzymes generated a patent. Another manuscript related to this technological extension was sent for review and under revision.
|
| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
The optimized single-molecule DNA recovery platform was successfully applied for high-throughput protein screening independent of cell culture. The same platform was successfully extended for use in other applications.
|
| Strategy for Future Research Activity |
I will push this study forward according to my initial plan and continue to explore the possibility of other extensions of my system in terms of analytical chemistry.
|
-
[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
-
Journal Title
Science Advances
Volume: 6
Pages: eaay0888
DOI
Peer Reviewed / Open Access
-
-
-
-
-
-
-
[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
-
-
-
-
-
-
