| Project/Area Number |
26650013
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Multi-year Fund |
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| Research Field |
Structural biochemistry
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| Research Institution | Hokkaido University |
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Principal Investigator |
YAO Min 北海道大学, 先端生命科学研究科(研究院), 教授 (40311518)
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| Co-Investigator(Renkei-kenkyūsha) |
UCHIUMI Toshio 新潟大学, 大学院自然科学研究科, 教授 (50143764)
KATO Koji 北海道大学, 大学院先端生命科学研究院, 助教 (30452428)
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| Project Period (FY) |
2014-04-01 – 2016-03-31
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| Project Status |
Completed (Fiscal Year 2015)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2015: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
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| Keywords | 翻訳速度 / 発現系 / ストーク複合体 / タンパク質工学 / リボソーム / X線結晶解析 |
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| Outline of Final Research Achievements |
The protein synthesis speed of ribosome is one of the important factors for over-expressing soluble protein using Escherichia coli, especially over-expressing eukaryotic proteins. In addition, it has been known that the bacterial protein synthesis speed (GTPases-turnover) is about 10 times of eukaryote. In order to construct an over-expression system with low speed of protein synthesis (translation) of E. coli, we tried to modify ribosomal stalk L10 from E. coli to a chimera L10 (L10P0) which binds to both ribosomal protein L12 and P1 (eukaryotic type), and has a chimera characters of bacteria and eukaryote of GTPases-turnover. We have successfully expressed mutants of L10ΔCH, L10ΔCH-P0H2CTD, and L10ΔCH-P0H3CTD. The binding assay of purified L10ΔCH-P0H2CTD with P1 showed the possibility for constructing a chimera stalk complex which will reduce the protein synthesis speed on ribosome.
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