| Project/Area Number |
61470150
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemical pharmacy
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| Research Institution | Osaka University |
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Principal Investigator |
UESUGI Seiichi Associate Professor Faculty of Pharmaceutical Sciences, Osaka University, 薬学部, 助教授 (70028851)
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| Co-Investigator(Kenkyū-buntansha) |
TANAKA Toshiki Protein Engineering Research Institute, 主任研究員 (70171775)
NISHIKAWA Satoshi Research Assistant Faculty of Pharmaceutical Sciences, Osaka University, 助手 (70150307)
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| Project Period (FY) |
1986 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1988: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1987: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1986: ¥5,000,000 (Direct Cost: ¥5,000,000)
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| Keywords | Phosphite method / H-Phosphonate / Ribooligonucleotide / solid-phase synthesis / RNA / phosphoramidite / NMR / ホスホアミダイト / 構造 / ホスホアミダイト体 / ホスホネート体 / オルトニトロベンジル基 / ホスホロアミダイト体 |
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| Research Abstract |
1. Synthesis of ribooligonucleotides using the phosphoramidite derivatives
Ribooligonucleotide derivatives (1), which are protected with an o-nitrobenzyl group at the 2'-hydroxyl, a methoxytrityl group at the 5'-hydroxyl and an acyl group at the base amino group, were synthesized. Methyl or cyanoethylidiisopropylamidite derivatives(2) were synthesized by phosohitylation at the 3'-hydroxyl group. On the other hand, nucleoside resin derivatives (3) were synthesized by coupling 1 with the controlled pore glass support. The oligomers were synthesized by repeated coupling of 2 with 3. When 5-(P-nitrophenyl)tetrazole is used for activation of the amidite, the coupling reaction is usually completed within one miunte.
2. Synthesis of ribooligonucleotides using the H-phosphonate derivatives
H-Phosphonate derivatives (4) were synthesized from 1 by treatment with PC1_3. The oligomers were synthesized by repeated coupling of 4 with 3 using pivaloyl chloride as the condensing agent. tRNA fragments with chain lengths up to 34 were synthesized by using a mucleic acid synthesizer.
3. Synthesis of ribooligonucleotides in quantity
A method for synthesis of the oligomers with chain lengths 10-20 by solid-phase phosphoramidite approach was established. Thus the RNA oligamers of abaout 100 absorbance units (260 nm), which can be used for NMR measurement, are easily synthesized in a short time in 10-20%yields.
4. Conformational studies on the ribooligonucleotides by NMR spectroscopy
Oligomer complexes having a catalytic activity or partial base sequence of an RNA enzyme and oligomers with extraordinary stable hairpin structures were synthesized by the above method and their conformation was examined by NMR.
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