Studies on the mechanisms of pre-mRNA splicing using synthetic oligonucleotides
| Project/Area Number |
61480464
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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 |
代謝生物化学
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| Research Institution | Kyushu University (1987)
University of Tsukuba (1986) |
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Principal Investigator |
OHSHIMA Yasumi Kyushu University, professor, 理学部, 教授 (90037606)
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| Co-Investigator(Kenkyū-buntansha) |
OHTSUKA Eiko Hokkaido University, professor, 薬学部, 教授 (80028836)
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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 |
¥6,000,000 (Direct Cost: ¥6,000,000)
Fiscal Year 1987: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1986: ¥4,000,000 (Direct Cost: ¥4,000,000)
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| Keywords | 2'-o-methylated oligonucleotides / Pre-mRNA splicing / Intron / Exon / Donor site / site / スキャニングモデル |
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| Research Abstract |
Seven kinds of 12mer 2'-o-methylated oligonucleotides were synthesized in the laboratory of E. Ohtsuka. Each of them are complementary to each of the seven portions in a pre-mRNA transcribed from the first exon-first inrton- second exon of human <beta>-globin gene. ANTIR-1 and 2 are complementary portions in the first exon. ANTIR-3 is complementary to the donor site. ANTIR-4 and 5 are complementary to portions within the intron. ANTIR-6 and 7 are complementary to portions of the second exon. The ^<32>P-labeled pre-mRNA was mixed with 1,000 fold excess of each of the oligos and incubated for hybridization. The mixbure was then tested for splicing activity in vitro using a HeLa cell nuclear extract. In a preliminary result, ANTIR-2 and 6 as well as 3 inhibited splicing of the pre-mRNA(mainly of the second step) while ANTIR-1, 4 and 5 did not. If the result is confirmed, it may be against a simple scanning model for splicing.
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Report
(2 results)
Research Products
(6 results)
