| Project/Area Number |
15K14228
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | Chiba University |
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Principal Investigator |
Umeno Daisuke 千葉大学, 大学院工学研究院, 准教授 (00400812)
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| Co-Investigator(Kenkyū-buntansha) |
河合 繁子 千葉大学, 大学院工学研究院, 助教 (40638920)
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| Project Period (FY) |
2015-04-01 – 2018-03-31
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| Project Status |
Completed (Fiscal Year 2017)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2016: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 合成生物学 / 制御 / 翻訳後修飾 / 進化分子工学 / プレニル化転移酵素 / イソプレノイド / カロテノイド / キナーゼ / スクリーニング / セレクション / プレニル化酵素 / 局在化 / 大腸菌 / バイオテクノロジー / 発現制御 / 生体機能利用 / 微生物 |
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| Outline of Final Research Achievements |
The ultimate purpose of this study is to reconstitute the protein-localization system via reconstitution of yeast prenylation enzymes Ram-1/-2 system in Escherichia coli. We succeeded in establishing the colorimetric screening method to indirectly visualize the activity of protein prenylation systems. Also established was the selection platform for protein prenylation by way of the monitoring of expression level of reporter genes under the control of transcription factors with prenylation tags. We also succeeded in the improvement of the selection system by elevating the efficacy of incorporating reagent called dP, the hyper-mutagenic nucleoside analogues, into genomic DNA thereby reducing the necessary concentration of dP added to the selection media. This way, we could have established the robust and reproducible selection system for improving protein prenylation systems in heterologous hosts.
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