| Project/Area Number |
07640936
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
系統・分類
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| Research Institution | The Nihon University |
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Principal Investigator |
INOUE Tadashi The Nihon Univ.College of Bioresorce Sciences, Professor, 生物資源科学部, 教授 (90124213)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1996: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
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| Keywords | Nosema bombycis / Ribosome / in vitro protein synthesis / 微胞子虫 / 試験管内蛋白質合成系 / リボソーム / 真核生物 |
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| Research Abstract |
In order to investigate the evolutionary origin of eukaryotes, we attempted to develop an in vitro protein synthesizing system consisting of ribosomes from some microsporidia including Nosema bombycis which is believed to diverge at early stage of eukaryotes. N.bombycis is known to have ribosomes whose sedimentation property is very similar to that of prokaryotes.
Crude cellular extract was prepared from both dormant and artificially hatched spores. Ribosomes was recovered from the extract by differential centrifugation and incubated with the soluble enzyme fraction in an in vitro protein synthesizing system. The system contained exogenously added poly (U), ^3H-Phe and an ATP-regenarating system. The amount of in vitro protein synthesis was measured by counting the radioactivity of hot trichloroacetic acid-insoluble fraction after incubation.
All the attempt to establish the system was failed because of poor activity of ribosomes and/or enzyme fraction of N.bombycis. Supplementation of the enzyme fraction of Ecsherichia coli, rat liver and midgut of silkworm had no effect on the activity. Aminoacyl-tRNA synthesis, however, took place normally in the system from N.bombycis. These results may indicate that some inhibitory factors are present in the preparation of N.bombycis ribosomes.
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