| Project/Area Number |
59480151
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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 |
Experimental pathology
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
NIWA Ohtsura (1986) Res. Inst. Nucl. Med. Biol., Hiroshima Univ.,Assoc. Prof., 原爆放射能医学研究所, 助教授 (80093293)
瀬山 敏雄 (1985) 広島大学, 原爆放射能医学研究所, 助手
濱田 勝友 (1984) 広島大学, 原爆放射能医学研究所, 助手
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| Co-Investigator(Kenkyū-buntansha) |
INOH Akira Res. Inst. Nucl. Med. Biol., Hiroshima Univ., Res. Assoc., 原爆放射能医学研究所, 助手 (30116909)
YOKORO Kenjiro Res. Inst. Nucl. Med. Biol., Hiroshima Univ., Prof., 原爆放射能医学研究所, 教授 (70034618)
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| Project Period (FY) |
1984 – 1986
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| Project Status |
Completed (Fiscal Year 1986)
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| Budget Amount *help |
¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 1986: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1985: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1984: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | Mouse thymoma / Transfection / Transformation / Transforming RNA / Mink lung cells |
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| Research Abstract |
This study is focused on in vitro cell transformation with small molecular weight RNA extracted from mouse thymoma cells. The RNA responsible for transforming activity was designated as transforming RNA. Total cellular RNA was obtained from chemically-induced lymphoma cells in mice by Guanidinium isothiocyanate (GTC)-hot phenol extraction. Transforming RNA was purified by the two cycles of electrophoresis through 20% native polyacrylamide gels or three cycles of electrophoresis through 10% denaturing polyacrylamide gels. The RNA was transfected into rat fibroblastic cell line, 3Y1 cells. When 3Y1 cells transfected with the RNA was maintained without serial passage, transformed cell foci appeared at the latency of 43 to 53 days. The transformed phenotype of 3Y1 cells was confirmed by colony formation of the cells in soft agar.
In order to confirm the purity of transforming RNA thus isolated, the RNA was sequenced by the direct chemical method using [3'-32P]RNA. The partial 110 bases sequence of the RNA was read. It was revealed that transforming RNA sequence had remarkable homology to one of the small nuclear RNA, namely U5 RNA.
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