| Project/Area Number |
16360260
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Civil and environmental engineering
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| Research Institution | Tohoku University |
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Principal Investigator |
OMURA Tatsuo Tohoku University, Graduate School of Engineering, Professor, 大学院工学研究科, 教授 (30111248)
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| Co-Investigator(Kenkyū-buntansha) |
WATANABE Toru Tohoku University, Graduate School of Engineering, Research Associate, 大学院工学研究科, 助手 (10302192)
佐野 大輔 東北大学, 大学院・工学研究科, 日本学術振興会特別研究員(PD)
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| Project Period (FY) |
2004 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥15,100,000 (Direct Cost: ¥15,100,000)
Fiscal Year 2006: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 2005: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 2004: ¥7,100,000 (Direct Cost: ¥7,100,000)
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| Keywords | Virus-Binding Proteins / Pathogenic viruses / Water environment / Immobilization / Polylysine-tag / ポリリジンタグ / PCR / ELISA / 配向固定化 / グルタルアルデヒド / ポリオウイルス / クローニング |
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| Research Abstract |
Virus-binding proteins (VBPs) are bacterial proteins to which human viruses preferentially adsorb in water environment. VBPs were discovered from a bacterial culture derived from activated sludge with affinity chromatography assay in which a viral capsid peptide was used as a ligand. The purpose of this study was to develop a novel technology using VBPs for virus removal from environmental water. Achievements described below were obtained during the 3-year study duration.
At the first year of the study duration, the scale-up of VBP production and purification was conducted in order to obtain large amount of VBP. A handmade column for VBP purification was constructed, which made it possible to obtain about 10 times larger volume of VBP than conventional procedure.
At the second year, gene of hexalysine-tagged VBP (LysTagVBP), which has 6-consecutive lysine residues in its C terminus, was fabricated with PCR, and used for production of LysTagVBP with E. coli. Then, polylysine region in LysTagVBP was coupled with the surface of glass particles using silane coupling reagent and glutaraldehyde. The creation of LysTagVBP directly linked to the effective immobilization of VBP on carrier surface.
At the third year of the study duration, the virus-removing column in which LysTagVBP was immobilized was constructed, and virus removal efficiency with the column was evaluated. As a result, about 40% of viruses were reduced during the first 1-hour operation, and 70% of viruses were removed after the 3-hour operation. The development of virus removal technology using VBPs is a totally new challenge, and the achievements with regards to the utilization of VBPs as viral adsorbent would be useful in the farther application of VBPs to virus removal from water.
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