| 研究課題/領域番号 |
21K06043
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| 研究機関 | 国立研究開発法人理化学研究所 |
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研究代表者 |
Malay Ali 国立研究開発法人理化学研究所, 環境資源科学研究センター, 上級研究員 (40467006)
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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 | silk / biomimetics / phase separation / spider / nanofibril / self-assembly / biopolymer |
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| 研究実績の概要 |
As outlined in the project proposal, I have designed and obtained recombinant genetic constructs for expression of spidroins in bacterial, with different domain and/or specific sequence combinations. Heterologous expression in E. coli was carried out, with highly promising results in the case of the MaSp2 spidroin constructs. For MaSp1, low levels of expression necessitated testing different fusion protein platforms for improved yields. Various initial experiments have been carried put to characterize the purified spidroin variants, including generation of phase separation maps, microscopy to probe mesoscale structure self-assembly , Raman spectroscopy, synchrotron small-angle X-ray scattering (SAXS), among others.
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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
So far, the project is progressing smoothly, and as planned. As mentioned above, in the case of the MaSp1 constructs, inherent issues with the protein solubility and overall yield might require further testing of different expression platforms using empirical methods.
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| 今後の研究の推進方策 |
From the second year of the project, I will focus on the self-assembly mechanisms and structures of the different spidroin constructs. Aside from experiments outlined in the original proposal, depending on the progress of the research, we may also explore alternative experimental approaches, such as super-resolution microscopy, NMR spectroscopy, microfluidics methods for self-assembly, and computational methods for modelling molecular interactions.
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| 次年度使用額が生じた理由 |
There was some delay in the experimental work and therefore the purchasing of some items was delayed. For the next year we will purchase consumable items that will make use of the extra funds.
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