| Project/Area Number |
61570988
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Chemical pharmacy
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| Research Institution | Hokkaido university |
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Principal Investigator |
IKEDA Kazuyoshi Center for Instrumental Analysis, Hokkaido Univ.,associate prof., 機器分析センター, 助教授 (20001042)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | deoxyoligonucleotides / putrescinylthymine / L-gultaminothymine / melting temperature / ヌクレアーゼ / 制限酵素 / DNA / 異常塩基 |
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| Research Abstract |
Oligonucleotides containing unusual base in w-14DNA,putrescinylthymine and its analogous bases were prepared by the usual method. Nucleosides,5-(6-hydroxyhexyl) and 5-(6-aminohexyl)-2'-deoxyuridines were prepared from 5-iodo-2'-deoxyuridine by the method of coupling reaction with hexyne derivative following by reduction. The synthetic method of the oligonucleotides containing 5-formyluracil base was developed by the modification of usual method, The reaction of formyl groups in their synthetic oligonucleotides with amines gave Schiff bases, which were reduced with sodium borohydride to obtain oligonucleotides containing 5-alkylaminomethyluracils, The reaction of 5-formyl-2'-deoxyuridine with L-gultanic acid following by the reduction gave 5-L-gultaminomethyl-2'-deoxyuridine which was unusual base in DNA of the bactriopharge sp-10 and PMB-1.
The duplex structure of their oligonucleotides was B-form conformation as determined with CD spectra. It was suggested that secondary amino group of 5-substituted function in their duplex oligonucleotides tends to decrease the thermal stabitity and the primary amino group increases it from melting temperature values. The thermal stability also depended on the sequence of oligonucleotide containing putrescinylthymine as which had GC pairing of adjacent to the unusual base showed large stability.
Synthetic oligonucleotides containing 5-alkylaminomethyluracil were resistant to some nucleases to form fragments with modified bases.
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