| Project/Area Number |
15K06585
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Multi-year Fund |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | University of Shizuoka |
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Principal Investigator |
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| Co-Investigator(Renkei-kenkyūsha) |
ITO Keisuke 静岡県立大学, 食品栄養科学部, 准教授 (40580460)
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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 |
¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000)
Fiscal Year 2017: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2016: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000)
Fiscal Year 2015: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
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| Keywords | protein expression / Recombinant protein / yeast expression system / protein synthesis / molecular biotechnology / bimolecular engineering / 遺伝子工学 / 難生産性蛋白質 / 菌体高密度化 / Ygr067C / Adr1 / 糖鎖修飾 / 組換え蛋白質生産 / 組換え酵母 / 菌体密度 / 遺伝子発現 / 分泌蛋白質 / 高度糖鎖修飾 |
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| Outline of Final Research Achievements |
Our novel expression system, namely high cell-density expression, have achieved successful expression of a certain range of “difficult-to-express” secretory protein while the molecular mechanisms behind it was remained to be studied. In this project, we identified two transcription factors, Adr1 and Ygr067C, as a potential host factor that is responsible for the cell-density-specific transcriptome change, where certain mRNAs of which transcriptional promoter shared typical Adr1p binding sequence. In fact, the yeast strain lacking Ygr067c showed a significant reduction in the cell-density dependent protein expression. The factor that directly allows the host to produce the difficult-to-express protein under the transcription factor is to be sought.
In parallel, we successfully find certain types of rare codons in recombinant gene help co-translational folding of synthesized protein, resulting in increased amount of recombinant protein in active form.
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