| 研究課題/領域番号 |
21K05341
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| 研究機関 | 京都大学 |
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研究代表者 |
Robert Martin 京都大学, 薬学研究科, 特定准教授 (90365487)
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| 研究期間 (年度) |
2021-04-01 – 2024-03-31
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| キーワード | biofilm / E. coli / cell motility / surface properties / nutrients / differentiation / gene expression |
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| 研究実績の概要 |
We focused our efforts on finding the conditions to generate reproducible biofilms with the expected structural complexity. This is expected to be important in providing an environment suitable for cellular differentiation and division of labor, one of the important focus of our study. By manipulating both the growth surface properties (agar hardness) and the nutritional conditions (salt-free agar with various concentrations of glucose) we produced biofilms having significant size and structural diversity. We also monitored the activity of several E. coli promoters using GFP reporter strains under these conditions. We observed patterns of promoter activity (gene expression) that varied widely depending on promoter and also as expected displayed some variation in spatial expression pattern.
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| 現在までの達成度 (区分) |
現在までの達成度 (区分)
2: おおむね順調に進展している
理由
The research is progressing as expected and we have been able to test multiple growth conditions and different promoters and E. coli strains. We are now ready the start the proteome analysis and continue the promoter activity analysis on a larger scale as originally planned.
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| 今後の研究の推進方策 |
We will continue to produce biofilms expressing different promoter reporters to monitor gene expression in space and time during development. This will be complemented with proteome analysis during the second year.
For gene expression analysis, in addition to the the E. coli fluorescent promoter collection we will also test and use a different set of YFP-fusion protein strains where gene expression can be directly visualized at the protein level.
To increase experimental throughput we will need extra sets of imaging systems, some of which are no longer commercially available. We are now planning to build some ourselves using open source resources and custom built and 3D-printed components.
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| 次年度使用額が生じた理由 |
Just at the beginning of the project the PI unexpectedly received an incubator from another researcher (transfer) delaying the need for the original purpose. When needed at the end of the fiscal year because of expansion of project scale, it was stock limitation and delivery delays made it impossible to secure in the first fiscal year. Also due to Covid-19 limitations, no business travel expenses were used. The unused money will be used early in the next fiscal year to purchase the incubator and other experimental resources and for travel if the conditions make it possible.
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