A Nobel cloning tool using the designed horizontal gene-transfer system by natural transformation
| Project/Area Number |
23710239
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| Research Category |
Grant-in-Aid for Young Scientists (B)
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| Allocation Type | Multi-year Fund |
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| Research Field |
Applied genomics
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
KANEKO Shinya 東京工業大学, 生命理工学研究科, 助教 (10399694)
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| Project Period (FY) |
2011 – 2013
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2011: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
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| Keywords | 応用微生物 / 核酸 / ゲノム / バイオテクノロジー / 形質転換 |
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| Research Abstract |
A designed horizontal gene-transfer system by natural transformation has been developed as novel cloning tool to introduce plasmid DNA into the objective cells without purification. Each culture condition for donor and recipient cells has been decided in this study. The bacteriolysis induction of donor cells by virulent phage is very important for universal use. Furthermore it was revealed that each aurintricarboxylic acid as nuclease inhibitor and exonuclease I as accelerator is effective for the DNA transfer by mixing it appropriately. In this condition it was found that the DNA transfer was extended to successful size increase up to 100 kb. The damage-free handling of DNA larger than 100 kb in test tubes is increasingly difficult. The designed transfer system named as Culture Mix Method (CMM) can apply various plasmid DNAs simply and conveniently, which should facilitate development of experimental tools to mobilize giant DNAs for genomic syntheses in synthetic biology.
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Report
(4 results)
Research Products
(6 results)
