| Project/Area Number |
24655145
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| Research Category |
Grant-in-Aid for Challenging Exploratory Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemistry related to living body
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| Research Institution | Hokkaido University |
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Principal Investigator |
TANAKA Yoshikazu 北海道大学, 先端生命科学研究科(研究院), 准教授 (20374225)
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| Project Period (FY) |
2012-04-01 – 2014-03-31
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| Project Status |
Completed (Fiscal Year 2013)
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| Budget Amount *help |
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2013: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2012: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000)
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| Keywords | tRNA / 精製 / 修飾 / 結晶化 / 結晶構造解析 / 結晶 |
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| Research Abstract |
In this study, I tried to establish conventional method to prepare highly pure modified tRNA from E. coli cell using tRNA-protein complex spontaneously formed in the cells. First, I established preparation system for tRNA(Met) using E. coli tRNA(Met) acetyltransferase. For this purpose, I selected K205A mutant, which has high affinity for tRNA. Moreover, crystals of the purified tRNA(Met) were obtained. The crystals diffracted about 6-angstrom resolution at synchrotron radiation facility. Furthermore, co-expression system of selenium introducing enzyme with tRNA(Gln), tRNA(Glu), and tRNA(Lys) in E. coli were constructed, and pure tRNAs were prepared from the tRNA-protein complex spontaneously formed in the E. coli cell.
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