Evolutionary design of protein prenyltion pathways

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K14228
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Biofunction/Bioprocess
• Research Institution: Chiba University
• Principal Investigator: Umeno Daisuke 千葉大学, 大学院工学研究院, 准教授 (00400812)
• Co-Investigator(Kenkyū-buntansha): 河合 繁子 千葉大学, 大学院工学研究院, 助教 (40638920)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 合成生物学 / 制御 / 翻訳後修飾 / 進化分子工学

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.

Free Research Field

進化合成生物学